Compounds for use as therapeutic agents affecting p53 expression and/or activity

ABSTRACT

The present disclosure relates to compound (I) 
     
       
         
         
             
             
         
       
     
     wherein R1 and R2 independently represent a hydrogen atom, a (C 1 -C 4 )alkoxy group, a fluoro(C 1 -C 4 )alkoxy group, a hydroxyl group, a benzyloxy group, a di(C 1 -C 4 )alkylamino group, a pyridyl-vinyl group, a pyrimidinyl-vinyl group, a styryl group, or a —NHCOphenyl group; R3, R4 and R5 independently represent a hydrogen atom, a (C 1 -C 4 )alkyl group, a CONHR6 group, a —CONR7R8 group, a —SO 2 NHR6 group, or a heteroaryl group optionally substituted by a halogen atom, a —(CH 2 ) n NR7R8 group or a hydroxy(C 1 -C 4 )alkyl group; R6 represents a hydrogen atom, a —(CHR9) m (CH 2 ) n NR7R8 group or a (C 1 -C 6 )alkyl group optionally substituted by a hydroxyl group; or anyone of its pharmaceutically acceptable salt, for use in a method for preventing, inhibiting or treating a disease in a patient suffering thereof, said disease involving a deregulated p53. Some of said compounds are new and also form part of the disclosure.

This is a Division of application Ser. No. 14/009,283 filed Oct. 1,2013, which in turn is a National Stage entry of PCT/IB12/51603 filedApr. 2, 2012, which claims the benefit of U.S. Provisional ApplicationNo. 61/470,564 filed Apr. 1, 2011, and European Patent Application No.11305385.4 filed on Apr. 1, 2011. The disclosure of the priorapplications is hereby incorporated by reference herein in its entirety.

FIELD OF THE INVENTION

The present invention is generally dedicated to compounds for use astherapeutic agents affecting p53 expression and/or activity.

BACKGROUND OF THE INVENTION

p53 is a house-keeping protein, the activity and expression of which, aswell as its cellular pathway, are critically implied in the maintainingof cell homeostasis, and the occurrence of numerous diseases. Indeed,numerous diseases may result from a deregulation of p53, either in itsexpression, or in its activity or cellular pathway, including itsactivating factors or its target genes.

The p53 gene is the most frequently mutated gene in cancers. Its tumoursuppressor activity relies on its ability to control proliferation byinducing apoptosis, cell cycle arrest and senescence in response tocellular stresses, such as DNA damages. Almost all human cancer exhibitdefects in p53 activation either by p53 gene mutation which occurs inaround 50% of human cancers, or by the failure (shortcoming) of thepathways promoting its activation.

The p53 pathway is also implicated in other diseases such asinflammation, fibrosis, neurodegenerative diseases, ischemia,atherosclerosis, pathogenesis of a number of hepatic disorders, such ascholestasis, autoimmune diseases, ethanol-induced injuries such asalcoholic liver diseases (ALD) including fatty liver, alcoholichepatitis and cirrhosis, ribosome biogenesis disorders such as TreacherCollins syndrome (TCS), male infertility, alopecia, neurologicaldefects, endocrinopathy syndrome (ANE syndrome), Shwachman-Diamondsyndrome (SDS) and neurofibromatosis type 1 (NF1), and HIV-associatedpathologies such as dementia, diabetes and myocardial infarction.

Indeed, in diseases other than cancer, the inhibition of wild-type p53may represent a new therapeutic strategy. Thus, inhibition of p53activity or cellular pathway might be highly beneficial in theprevention of injury to diverse organs (Georgiev et al., Curr Pharm Des,2006 12:2911), or in treatment of myocardial infarction (Matsusaka etal., 2006). Also, ischemia, or diseases associated withneurodegenerative pathological conditions, including AIDS-associatedneurodegeneration, stroke, Parkinson's, Alzheimer's, and Huntington'sdiseases (Vousden & Lane, 2007, Nat Rev Mol Cell Biol. 2007 Apr.;8(4):275-83), clinical management of hepatic disease induced by toxicbile salts (Oh et al., 2002 Toxicol Appl Pharmacol. 2002 Jul. 1;182(1):27-33), or atherosclerosis in which wild-type p53 isover-expressed (Iacopetta et al., Int J Oncol, 1995, 7, 399-402), maybenefit from compounds able to curb deregulated p53.

Document WO2009/087238 discloses compounds effective in treatingdiseases related to the splicing process.

Up to now, to the knowledge of the inventors, no drug repressing mutatedp53 expression has been described. Indeed activation of p53 pathway isgenerally obtained but no reduction of p53 pathway.

Therefore, there is an existing need to find out compounds able tocompensate deregulated p53 expression, activity, or cellular pathway.

More particularly, there is a need to identify new compounds able toinhibit or reduce the expression of mutated p53 isoforms, preferablywithout affecting the expression of wild-type p53 isoforms.

There is a need to have compounds able to favour the expression ofnon-mutated p53 isoforms over mutated p53 isoforms.

There is also a need to have compounds able to modulate the p53 geneexpression, and in particular able to specifically modulate the splicingof p53 isoforms.

There is also a need to have compounds able to prevent or reduce overexpression of wild-type isoform(s) of p53.

There is also a need to have compounds able to prevent or increaselow-expression of wild-type isoform(s) of p53.

More precisely, there is a need to find out compounds able to prevent,inhibit and/or treat a disease in a patient suffering thereof, saiddisease involving deregulated p53.

More particularly, there is a need to have compounds able to prevent,inhibit and/or treat a disease in a patient suffering thereof, saiddisease involving mutated p53, and/or deregulated wild-type p53.

More particularly, there is a need to have compounds able to prevent,inhibit and/or treat a cancer in a patient suffering thereof, saidcancer involving a mutated p53 or deregulated wild-type p53, such as alow-expression of wild-type p53.

There is a need to have compounds able to prevent, inhibit and/or treatatherosclerosis in a patient suffering thereof, said atherosclerosisinvolving a mutated p53, or a deregulated wild-type isoform of p53, suchas a low-expression of wild-type p53.

There is also a need to have compounds able to prevent, inhibit and/ortreat a disease in a patient suffering thereof, said disease involving aderegulated wild-type p53, such as a disease chosen from inflammation,fibrosis, neurodegenerative diseases, ischemia, atherosclerosis, hepaticdisorders, such as cholestasis, autoimmune diseases, and ethanol-inducedinjuries such as alcoholic liver diseases (ALD) including fatty liver,alcoholic hepatitis and cirrhosis, ribosome biogenesis disorders such asTreacher Collins syndrome (TCS), male infertility, alopecia,neurological defects, endocrinopathy syndrome (ANE syndrome),Shwachman-Diamond syndrome (SDS) and neurofibromatosis type 1 (NF1), andHIV-associated pathologies such as dementia, diabetes and myocardialinfarction.

SUMMARY OF THE INVENTION

It has now been found that derivatives of formula (I) as definedhereinafter, which are partially described in document WO 2009/087238,are able to modulate the p53 expression and/or activity with adiscriminating effect over wild-type and mutated p53, as well as atisoforms level, as illustrated in the experimental data hereinafter.

More particularly, said derivatives are able to inhibit the expressionof mutated p53 and/or to modulate the expression of wild-type p53.Through their ability to modulate the expression of mutated p53 andpossibly wild-type p53, specifically at an isoform level, i.e. short orlong, the compounds of the invention are also advantageously able tomodulate the activity or cellular pathway of those proteins, and inparticular to affect the p53-dependent expression of target genes, suchas Bcl2, Hdm2 or p21. Accordingly, the derivatives of the invention areable to curb a deregulated p53 expression, activity or cellular pathway.

The derivatives of formula (I) as described hereinafter are moreoverable to exert a specific action on the short (truncated) or long isoformof p53.

Said compounds can selectively act on tumours involving a mutated p53.In other words, said compounds are particularly useful for treatingpatients affected by cancer and/or atherosclerosis involving a mutatedp53, and more particularly expression of a mutated p53.

Also, the compounds of the invention are able to curb the expression,activity or cellular pathway of wild-type p53, more particularly incells where wild-type p53 is over-expressed.

On the basis of such activity, the compounds are useful in theprevention, inhibiting and/or treatment of a disease involving aderegulated p53, such as a mutated p53 or a deregulated wild-type p53'spathway. In particular, the compounds of the invention are useful forpreventing and/or treating cancer and/or atherosclerosis involving amutated p53, and/or a deregulated wild-type p53, as well as forpreventing, inhibiting and/or treating diseases involving a deregulatedwild-type p53, such as inflammation, fibrosis, neurodegenerativediseases, ischemia, atherosclerosis, pathogenesis of a number of hepaticdisorders, such as cholestasis, autoimmune diseases, and ethanol-inducedinjuries such as alcoholic liver diseases (ALD) including fatty liver,alcoholic hepatitis and cirrhosis, ribosome biogenesis disorders such asTreacher Collins syndrome (TCS), male infertility, alopecia,neurological defects, endocrinopathy syndrome (ANE syndrome),Shwachman-Diamond syndrome (SDS) and neurofibromatosis type 1 (NF1), andHIV-associated pathologies such as dementia, diabetes and myocardialinfarction.

The compounds of the invention may be useful in the prevention and/ortreatment of cancer involving a mutated p53.

More particularly the compounds are useful in the prevention andtreatment of primary breast cancer, axillary node metastasis, HER2positive and ER negative cancers with poor prognosis, and diseasesinvolving a mutated p53, and in particular involving Δ133p53β isoform,and more particularly breast cancer and colon cancer.

The present invention therefore relates to compounds of formula (I) asdefined below for use in a method for preventing, inhibiting or treatingcancer and/or atherosclerosis in patients expressing a mutated p53, orinflammation, fibrosis, neurodegenerative diseases, ischemia,atherosclerosis, pathogenesis of a number of hepatic disorders such ascholestasis, autoimmune diseases and ethanol-induced injuries such asalcoholic liver diseases (ALD) including fatty liver, alcoholichepatitis and cirrhosis, ribosome biogenesis disorders such as TreacherCollins syndrome (TCS), male infertility, alopecia, neurologicaldefects, endocrinopathy syndrome (ANE syndrome), Shwachman-Diamondsyndrome (SDS) and neurofibromatosis type 1 (NF1), and HIV-associatedpathologies such as dementia, diabetes and myocardial infarction inpatients in which wild-type of p53 is deregulated.

The neurodegenerative diseases more particularly considered may bechosen from AIDS-associated neurodegeneration, stroke, Parkinson's,Alzheimer's, and Huntington's diseases.

The present invention moreover relates to a method of preventing,inhibiting or treating diseases involving a deregulated p53, inparticular a mutated p53 or a deregulated wild-type p53.

More particularly, the invention relates to a method for preventing,inhibiting or treating a cancer in patients said cancer involving amutated p53, or a disease chosen from fibrosis, neurodegenerativediseases, ischemia, cholestasis or atherosclerosis in patients, saiddisease involving deregulation of expression, activity or pathway ofwild-type p53, which comprises at least one step consisting inadministering to a patient suffering thereof an effective amount of acompound as defined in formula (I) below or one of its pharmaceuticallyacceptable salts.

The present invention further relates to some particular derivatives assuch, as defined below.

The present invention also provides pharmaceutical compositionscomprising at least one of said particular compounds.

DETAILED DESCRIPTION OF THE INVENTION

According to a first aspect, a subject-matter of the present inventionrelates to a compound of formula (I)

wherein:

R1 and R2 independently represent

-   -   a hydrogen atom, a (C₁-C₄)alkoxy group, a fluoro(C₁-C₄)alkoxy        group, a hydroxyl group, a benzyloxy group, a        di(C₁-C₄)alkylamino group,    -   a pyridyl-vinyl or pyrimidinyl-vinyl group, said two groups        being optionally substituted by a halogen atom, a (C₁-C₄)alkyl        group or a fluoro(C₁-C₄)alkyl group,    -   a styryl group optionally substituted by a (C₁-C₄)alkoxy group        or a fluoro(C₁-C₄)alkoxy group,    -   a —NHCOphenyl group, the phenyl group being optionally        substituted by a (C₁-C₄)alkoxy group or a fluoro(C₁-C₄)alkoxy        group,

R3, R4 and R5 independently represent

-   -   a hydrogen atom, a (C₁-C₄)alkyl group,    -   a —CONHR6 group,    -   a —CONR7R8 group,    -   a —SO₂NHR6 group,    -   a heteroaryl group optionally substituted by a halogen atom, a        —(CH₂)_(n)NR7R8 group or a hydroxy(C₁-C₄)alkyl group,

R6 represents a hydrogen atom, a —(CHR9)_(m)(CH₂)_(n)NR7R8 group or a(C₁-C₆)alkyl group optionally substituted by a hydroxyl group,

R7 and R8 independently represent a (C₁-C₄)alkyl group or form togetherwith the nitrogen atom to which they are attached a saturated orinsaturated 5- or 6-membered ring optionally containing a furtherheteroatom chosen among nitrogen and oxygen, said ring being optionallysubstituted by a (C₁-C₄)alkyl group,

R9 represents a (C₁-C₂)alkyl group,

m is 0 or 1,

n is 1, 2, 3, 4 or 5,

provided that two of R3, R4 and R5 are a hydrogen atom or a (C₁-C₄)alkylgroup and the last is different from a hydrogen atom or a (C₁-C₄)alkylgroup, and

or anyone of its pharmaceutically acceptable salt,

for use in a method for preventing, inhibiting or treating a disease ina patient suffering thereof, said disease involving a deregulated p53.

Within the invention, “a disease involving a deregulated p53” intends toencompass diseases where at least one isoform of p53, α, β or γ,full-length or truncated (short), has its expression, activity orcellular pathway altered, namely suppressed, reduced or increased.

The expression of p53 may be altered at a gene, mRNA and/or proteinlevel.

The expression and/or activity and/or cellular pathway of p53 may bealtered by a presence of a mutation in p53 or by an alteration of itsactivating factors.

The activity or the cellular pathway of p53 may be assessed bydetermination of the expression level of downstream target genes, suchas Hdm-2, p21, Bcl-2 or Bax.

In one embodiment, a disease involving a deregulated p53 may be chosenfrom a disease involving a mutated p53 or a disease involving aderegulated wild-type p53.

Within the invention, “mutated p53” intends to encompass mutation in thep53 gene resulting either in a decrease or absence of p53 proteinexpression, or in an increase of p53 protein expression, or inexpression of a mutated p53 protein exhibiting reduced, suppressed orincreased wild-type activity. The alteration of expression and/oractivity of mutated p53 may affect the whole family of p53, or only oneisoform, or at least one isoform. Preferably, a mutated p53 relates toat least one mutated isoform of p53, such as an isoform chosen from α, βor γ, truncated or full-length.

Preferably, the invention relates to diseases involving mutated p53. Inparticular, diseases involving mutated p53 exhibit reduced or suppressedactivity and/or expression of the mutated p53. More particularly suchdiseases may be chosen from cancer and/or atherosclerosis. In oneembodiment, the invention relates more particularly to a cancerinvolving at least one mutated isoform of p53, preferably Δ133p53, andin particular a β and/or γ isoform(s).

According to another embodiment, the invention relates to diseasesinvolving a deregulated wild-type p53. A “deregulated wild-type p53”encompasses suppressed, reduced or increased expression of wild-typep53, or suppressed, reduced or increased activity or cellular pathway ofwild-type p53. “Wild-type p53” intends to relate to at least one isoformof p53, such as chosen from α, β or γ, truncated or full-length.

Preferably, diseases involving a deregulated wild-type p53 may be chosenfrom inflammation, fibrosis, neurodegenerative diseases, ischemia,atherosclerosis, hepatic disorders, such as cholestasis, autoimmunediseases, ethanol-induced injuries such as alcoholic liver diseases(ALD) including fatty liver, alcoholic hepatitis and cirrhosis, ribosomebiogenesis disorders such as Treacher Collins syndrome (TCS), maleinfertility, alopecia, neurological defects, endocrinopathy syndrome(ANE syndrome), Shwachman-Diamond syndrome (SDS) and neurofibromatosistype 1 (NF1), and HIV-associated pathologies such as dementia, diabetesand myocardial infarction.

In one embodiment, the instant invention relates to a compound offormula (I) for use in a method for preventing, inhibiting or treatingcancer, inflammation, fibrosis, neurodegenerative diseases, ischemia,atherosclerosis, hepatic disorders, cholestasis, atherosclerosis,autoimmune diseases, ethanol-induced injuries such as alcoholic liverdiseases (ALD) including fatty liver, alcoholic hepatitis and cirrhosis,ribosome biogenesis disorders such as Treacher Collins syndrome (TCS),male infertility, alopecia, neurological defects, endocrinopathysyndrome (ANE syndrome), Shwachman-Diamond syndrome (SDS) andneurofibromatosis type 1 (NF1), and HIV-associated pathologies such asdementia, diabetes and myocardial infarction.

In a preferred embodiment, the invention relates to a compound offormula (I) as above-defined as an agent for preventing, inhibiting ortreating a cancer in a patient exhibiting a mutated p53, or fibrosis,neurodegenerative diseases, ischemia, cholestasis or atherosclerosis ina patient exhibiting a deregulated wild-type p53.

The term “preventing”, as used herein, means reducing the risk of onsetor slowing the occurrence of a given phenomenom, namely in the presentinvention, a disease involving a deregulated p53 such as cancer,inflammation, fibrosis, neurodegenerative diseases, ischemia,atherosclerosis, hepatic disorders, cholestasis, atherosclerosis,autoimmune diseases, ethanol-induced injuries such as alcoholic liverdiseases (ALD) including fatty liver, alcoholic hepatitis and cirrhosis,ribosome biogenesis disorders such as Treacher Collins syndrome (TCS),male infertility, alopecia, neurological defects, endocrinopathysyndrome (ANE syndrome), Shwachman-Diamond syndrome (SDS) andneurofibromatosis type 1 (NF1), and HIV-associated pathologies such asdementia, diabetes and myocardial infarction.

The pyridyl-vinyl group can be a 4-pyridyl-vinyl group, a3-pyridyl-vinyl group or a 2-pyridyl-vinyl group and preferably a4-pyridyl-vinyl group or a 2-pyridyl-vinyl.

When R7 and R8 form together with the nitrogen atom to which they areattached a saturated or insaturated 5- or 6-membered ring, said ring maypreferably be a imidazolyl, a morpholinyl, a piperidinyl, a pyrrolidinylor 4-methylpiperazinyl group.

According to one embodiment, the present invention relates to a compoundof formula (I) as defined above, wherein one of R1 and R2 is a hydrogenatom and the other is different from a hydrogen atom, for use in amethod for preventing, inhibiting or treating a disease in a patientsuffering thereof, said disease involving deregulated p53.

According to another embodiment, the present invention relates to acompound of formula (I) as defined above, wherein R3 represents ahydrogen atom and one of R4 and R5 is a —CONHR6 group or a heteroarylgroup optionally substituted by a —(CH₂)_(n)NR7R8 group or ahydroxy(C₁-C₄)alkyl group, wherein R6, R7, R8, R9 and n are as definedabove, for use in a method for preventing, inhibiting or treatingdisease in a patient suffering thereof, said disease involvingderegulated p53.

According to another embodiment, the present invention relates to acompound of formula (I)

wherein:

R1 and R2 independently represent

-   -   a hydrogen atom, a methoxy group, a trifluoromethoxy group, a        hydroxyl group, a benzyloxy group, a dimethylamino group,    -   a pyridyl-vinyl or pyrimidinyl-vinyl group, said two groups        being optionally substituted by a halogen atom, a methoxy group        or a trifluoromethyl group,    -   a styryl group optionally substituted by a methoxy group or a        trifluoromethoxy group,    -   a —NHCOphenyl group, the phenyl group being optionally        substituted by a methoxy group or a trifluoromethoxy group,

R3, R4 and R5 independently represent

-   -   a hydrogen atom, a methyl group,    -   a —CONHR6 group,    -   a —CONR7R8 group,    -   a —SO₂NHR6 group, or    -   a triazolyl or imidazolyl group, said group being optionally        substituted by a chlorine atom, a —(CH₂)_(n)NR7R8 group or a        hydroxy(C₁-C₄)alkyl group,

R6 represents a hydrogen atom, a —(CHR9)_(m)(CH₂)_(n)NR7R8 group or a(C₁-C₆)alkyl group optionally substituted by a hydroxyl group,

R7 and R8 independently represent a (C₁-C₄)alkyl group or form togetherwith the nitrogen atom to which they are attached a saturated orinsaturated 5- or 6-membered ring optionally containing a furtherheteroatom chosen among nitrogen and oxygen, said ring being optionallysubstituted by a methyl group, and said ring being chosen among aimidazolyl, a morpholinyl, a piperidinyl, a pyrrolidinyl or4-methylpiperazinyl group,

R9 represents a (C₁-C₂)alkyl group,

m is 0 or 1,

n is 1, 2 or 3,

provided that two of R3, R4 and R5 are a hydrogen atom or a methyl groupand the last is different from a hydrogen atom or a methyl group andthat one of R1 and R2 is a hydrogen atom and the other is different froma hydrogen atom, and

or anyone of its pharmaceutically acceptable salt,

for use in a method for preventing, inhibiting or treating a disease ina patient suffering thereof, said disease involving deregulated p53.

The compound of formula (I) in any embodiment as described above is moreparticularly useful as an agent for preventing, inhibiting or treating adisease in a patient, wherein the disease involves mutated p53 oraltered wild-type p53 expression, activity or pathway.

In the context of the present invention, the term “patients sufferingfrom diseases involving a mutated p53 gene or an altered wild-type p53expression, activity or pathway” means a specific group of patients forwhich it has been detected by known methods, for example sequencing orimmunodetection of mutated or wild-type p53 isoform(s) or methods aimingat assessing the expression, activity or pathway of p53, such as theassessment of p53 targets (Bcl-2, Hdm-2 or p21 . . . ), that p53mutation(s) exists or a mutated p53 is expressed or that the expression,activity or cellular pathway of wild-type p53 is altered.

The compound of formula (I) in any embodiment as described above is moreparticularly useful as an agent for preventing, inhibiting or treating adisease in a patient wherein the disease involves an altered expressionof wild-type p53, such as a low- or an over-expression, and inparticular an over-expression.

The compounds of the invention may exist in the form of free bases or ofaddition salts with pharmaceutically acceptable acids.

Suitable physiologically acceptable acid addition salts of compounds offormula (I) include hydrobromide, tartrate, citrate, trifluoroacetate,ascorbate, hydrochloride, tartrate, triflate, maleate, mesylate,formate, acetate and fumarate.

The compounds of formula (I) and or salts thereof may form solvates(e.g. hydrates) and the invention includes all such solvates.

The terms “hydrates” and “solvates” simply mean that the compounds (I)according to the invention can be in the form of a hydrate or a solvate,i.e. combined or associated with one or more water or solvent molecules.This is only a chemical characteristic of such compounds, which can beapplied for all organic compounds of this type.

In the context of the present invention, the term:

-   -   “halogen” is understood to mean chlorine, fluorine, bromine, or        iodine, and in particular denotes chlorine, fluorine or bromine,    -   “(C₁-C₄)alkyl” as used herein respectively refers to C₁-C₄        normal, secondary or tertiary saturated hydrocarbon. Examples        are, but are not limited to, methyl, ethyl, 1-propyl, 2-propyl,    -   “(C₁-C₄)alkoxy” as used herein respectively refers to        —O—(C₁-C₄)alkyl moiety, wherein alkyl is as defined above.        Examples are, but are not limited to, methoxy, ethoxy,        1-propoxy, 2-propoxy,    -   “fluoroalkyl group” and “fluoroalkoxy group” refers respectively        to alkyl group and alkoxy group as above-defined, said groups        being substituted by at least one fluorine atom. Examples are        perfluoroalkyl groups, such as trifluoromethyl, or        perfluoropropyl or perfluoroalkoxy groups, such as        trifluoromethoxy,    -   “heteroaryl” group refers to an aromatic ring, where the ring        structure is formed by 3 to 5 carbon atoms and by one to three        hetereoatoms chosen among N, O or S. When the heteroaryl group        is substituted, said heteroaryl group further bears one or more        substituents. The following heteroaryl may be cited: thienyl,        furyl, pyrrolyl, imidazolyl, pyrazolyl, pyridyl, pyrazinyl,        pyrimidinyl, pyridazinyl, triazolyl, and preferably triazolyl        and imidazolyl, and    -   “patient” may extend to humans or domestic mammals or mammals of        any economic value, such as cats or dogs.

According to a particular embodiment, an additional subject-matter ofthe present invention is a compound of formula (Iab)

wherein:

X, Y and Z are independently C or N,

R3, R4 and R5 are as defined above in anyone of the embodiments offormula (I),

provided that the lateral group

is in the meta or para position with respect to the —NH— group, andpreferably in the meta position,

provided that at most two of X, Y and Z are N, and that when Y is N, Xand Z are C, and

provided that two of R3, R4 and R5 are a hydrogen atom or a (C₁-C₄)alkylgroup and the last is different from a hydrogen atom or a (C₁-C₄)alkylgroup,

or one of its pharmaceutically acceptable salt,

for use in a method for preventing, inhibiting or treating a disease ina patient suffering thereof, said disease involving deregulated p53.

According to a more particular aspect of said embodiment, an additionalsubject-matter of the present invention is a compound of formula (Iab)

wherein:

R3 is hydrogen or a methyl group,

one of R4 and R5 represents

-   -   a —CONHR6 group, or    -   a triazolyl group being optionally substituted by a        —(CH₂)_(n)NR7R8 group or a hydroxy(C₁-C₄)alkyl group,

R6 represents a (C₁-C₆)alkyl group or a —(CH₂)_(n)NR7R8 group,

R7 and R8 independently represent a (C₁-C₄)alkyl group or form togetherwith the nitrogen atom to which they are attached a saturated orinsaturated 5- or 6-membered ring optionally containing a furtherheteroatom chosen among nitrogen and oxygen, said ring being optionallysubstituted by a methyl group, and said ring being chosen among aimidazolyl, a morpholinyl, a piperidinyl, a pyrrolidinyl or4-methylpiperazinyl group,

n is 1, 2 or 3,

and the other of R4 and R5 is a hydrogen atom,

or one of its pharmaceutically acceptable salt,

for use in a method for preventing, inhibiting or treating a disease ina patient suffering thereof said disease involving deregulated p53.

According to another particular embodiment, an additional subject-matterof the present invention is a compound of formula (Ia)

wherein:

R3, R4 and R5 are as defined above in anyone of the embodiments offormula (I),

provided that the lateral group

is in the meta or para position with respect to the —NH— group, andpreferably in the meta position, and

provided that two of R3, R4 and R5 are a hydrogen atom or a (C₁-C₄)alkylgroup and the last is different from a hydrogen atom or a (C₁-C₄)alkylgroup,

or one of its pharmaceutically acceptable salt,

for use in a method for preventing, inhibiting or treating a disease ina patient suffering thereof, said disease involving deregulated p53.

According to a more particular aspect of said embodiment, an additionalsubject-matter of the present invention is a compound of formula (Ia)

wherein:

R3 is hydrogen or a methyl group,

one of R4 and R5 represents

-   -   a —CONHR6 group    -   a SO₂NHR6 group,    -   a CONR7R8 group, or    -   a triazolyl group being optionally substituted by a        —(CH₂)_(n)NR7R8 group or a hydroxy(C₁-C₄)alkyl group,

R6 represents a hydrogen atom, a (C₁-C₆)alkyl group or a —(CH₂)_(n)NR7R8group,

R7 and R8 independently represent a (C₁-C₄)alkyl group or form togetherwith the nitrogen atom to which they are attached a saturated orinsaturated 5- or 6-membered ring optionally containing a furtherheteroatom chosen among nitrogen and oxygen, said ring being optionallysubstituted by a methyl group, and said ring being chosen among aimidazolyl, a morpholinyl, a piperidinyl, a pyrrolidinyl or4-methylpiperazinyl group,

n is 1, 2 or 3,

and the other of R4 and R5 is a hydrogen atom,

or one of its pharmaceutically acceptable salt,

for use in a method for preventing, inhibiting or treating a disease ina patient suffering thereof, said disease involving a deregulated p53.

According to another particular embodiment, an additional subject-matterof the present invention is a compound of formula (Ib)

wherein:

R3, R4 and R5 are as defined above in anyone of the embodiments offormula (I),

provided that the lateral group

is in the meta or para position with respect to the —NH— group, andpreferably in the meta position, and

provided that two of R3, R4 and R5 are a hydrogen atom or a (C₁-C₄)alkylgroup and the last is different from a hydrogen atom or a (C₁-C₄)alkylgroup,

or one of its pharmaceutically acceptable salt,

for use in a method for preventing, inhibiting or treating a disease ina patient suffering thereof, said disease involving a deregulated p53.

According to a more particular aspect of said embodiment, an additionalsubject-matter of the present invention is a compound of formula (Ib)

wherein:

R3 is hydrogen or a methyl group, one of R4 and R5 represents

-   -   a —CONHR6 group, or    -   a triazolyl group being optionally substituted by a        —(CH₂)_(n)NR7R8 group or a hydroxy(C₁-C₄)alkyl group,

R6 represents a (C₁-C₆)alkyl group or a —(CH₂)_(n)NR7R8 group,

R7 and R8 independently represent a (C₁-C₄)alkyl group or form togetherwith the nitrogen atom to which they are attached a saturated orinsaturated 5- or 6-membered ring optionally containing a furtherheteroatom chosen among nitrogen and oxygen, said ring being optionallysubstituted by a methyl group, and said ring being chosen among aimidazolyl, a morpholinyl, a piperidinyl, a pyrrolidinyl or4-methylpiperazinyl group,

n is 1, 2 or 3,

and the other of R4 and R5 is a hydrogen atom,

or one of its pharmaceutically acceptable salt,

for use in a method for preventing, inhibiting or treating a disease ina patient suffering thereof, said disease involving a deregulated p53.

According to another particular embodiment, an additional subject-matterof the present invention is a compound of formula (Ic)

wherein:

R10 is a hydrogen atom, a (C₁-C₄)alkoxy group or a fluoro(C₁-C₄)alkoxygroup,

R3, R4 and R5 are as defined above in anyone of the embodiments offormula (I),

provided that two of R3, R4 and R5 are a hydrogen atom or a (C₁-C₄)alkylgroup and the last is different from a hydrogen atom or a (C₁-C₄)alkylgroup,

or one of its pharmaceutically acceptable salt,

for use in a method for preventing, inhibiting or treating a disease ina patient suffering thereof, said disease involving a deregulated p53.

According to a more particular aspect of said embodiment, an additionalsubject-matter of the present invention is a compound of formula (Ic)

wherein:

R10 is a hydrogen atom, a methoxy group or a trifluoromethoxy group,

R3 is hydrogen or a methyl group,

one of R4 and R5 represents a —CONHR6 group,

-   -   R6 represents a (C₁-C₆)alkyl group or a —(CH₂)_(n)NR7R8 group,    -   R7 and R8 independently represent a (C₁-C₄)alkyl group or form        together with the nitrogen atom to which they are attached a        saturated or insaturated 5- or 6-membered ring optionally        containing a further heteroatom chosen among nitrogen and        oxygen, said ring being optionally substituted by a methyl        group, and said ring being chosen among a imidazolyl, a        morpholinyl, a piperidinyl, a pyrrolidinyl or        4-methylpiperazinyl group,    -   n is 1, 2 or 3,

and the other of R4 and R5 is a hydrogen atom,

or one of its pharmaceutically acceptable salt,

for use in a method for preventing, inhibiting or treating diseases adisease in a patient suffering thereof, said disease involving aderegulated p53.

According to another particular embodiment, an additional subject-matterof the present invention is a compound of formula (Id)

wherein:

R1, R2 and R6 are as defined above in anyone of the embodiments offormula (I),

R4 and R5 are independently a hydrogen atom or a (C₁-C₄)alkyl group,

or one of its pharmaceutically acceptable salt,

for use in a method for preventing, inhibiting or treating a disease ina patient suffering thereof, said disease involving a deregulated p53.

According to a more particular aspect of said embodiment, an additionalsubject-matter of the present invention is a compound of formula (Id)

wherein:

R1 and R2 independently represent a hydrogen atom, a methoxy group, atrifluoromethoxy group, a hydroxyl group, a benzyloxy group or adimethylamino group,

R6 represents a (C₁-C₆)alkyl group or a —(CHR9)_(m)(CH₂)_(n)NR7R8 group,

R7 and R8 independently represent a (C₁-C₄)alkyl group or form togetherwith the nitrogen atom to which they are attached a saturated orinsaturated 5- or 6-membered ring optionally containing a furtherheteroatom chosen among nitrogen and oxygen, said ring being optionallysubstituted by a methyl group, and said ring being chosen among aimidazolyl, a morpholinyl, a piperidinyl, a pyrrolidinyl or4-methylpiperazinyl group,

R9 represents a (C₁-C₂)alkyl group,

m is 0 or 1,

n is 1, 2 or 3,

provided that one of R1 and R2 is a hydrogen atom and the other isdifferent from a hydrogen atom,

or anyone of its pharmaceutically acceptable salt,

for use in a method for preventing, inhibiting or treating a disease ina patient suffering thereof, said disease involving a deregulated p53.

According to another particular embodiment, an additional subject-matterof the present invention is a compound of formula (Ie)

wherein:

R3 is a hydrogen atom or a (C₁-C₄)alkyl group,

R5 is as defined above in anyone of the embodiments of formula (I), and

R12 is a hydrogen atom, a (C₁-C₄)alkoxy group or atrifluoro(C₁-C₄)alkoxy group,

or one of its pharmaceutically acceptable salt,

for use in a method for preventing, inhibiting or treating a disease ina patient suffering thereof, said disease involving a deregulated p53.

According to a more particular aspect of said embodiment, an additionalsubject-matter of the present invention is a compound of formula (Ie)

wherein:

R3 is a hydrogen atom or a methyl group,

R5 represents

-   -   a —CONHR6 group, or    -   a triazolyl or imidazolyl group, said groups being optionally        substituted by a chlorine atom, a —(CH₂)_(n)NR7R8 group or a        hydroxy(C₁-C₄)alkyl group,

R6 represents a (C₁-C₆)alkyl group or a —(CH₂)_(n)NR7R8 group,

R7 and R8 independently represent a (C₁-C₄)alkyl group or form togetherwith the nitrogen atom to which they are attached a saturated orinsaturated 5- or 6-membered ring optionally containing a furtherheteroatom chosen among nitrogen and oxygen, said ring being optionallysubstituted by a methyl group, and said ring being chosen among aimidazolyl, a morpholinyl, a piperidinyl, a pyrrolidinyl or4-methylpiperazinyl group,

n is 1, 2 or 3,

R12 is a hydrogen atom, a methoxy group, a trifluoromethoxy group or abenzyloxy group,

or one of its pharmaceutically acceptable salt,

for use in a method for preventing, inhibiting or treating a disease ina patient suffering thereof, said disease involving a deregulated p53.

According to another particular embodiment, an additional subject-matterof the present invention is a compound of formula (If)

wherein:

R3 is a hydrogen atom or a (C₁-C₄)alkyl group,

R13 is a hydrogen atom, a (C₁-C₄)alkoxy group or atrifluoro(C₁-C₄)alkoxy group and can be in ortho, meta or para position,

R14, which can be in meta or para position, represents

-   -   a —CONHR6 group,    -   a heteroaryl group optionally substituted by a —(CH₂)_(n)NR7R8        group or a hydroxy(C₁-C₄)alkyl group,

R6 represents a (C₁-C₆)alkyl group optionally substituted by a hydroxylgroup,

or a —(CH₂)_(n)NR7R8 group,

R7 and R8 independently represent a (C₁-C₄)alkyl group or form togetherwith the nitrogen atom to which they are attached a saturated orinsaturated 5- or 6-membered ring optionally containing a furtherheteroatom chosen among nitrogen and oxygen and optionally substitutedby a (C₁-C₄)alkyl group, and said ring being chosen among a imidazolyl,a morpholinyl, a piperidinyl, a pyrrolidinyl or 4-methylpiperazinylgroup,

or one of its pharmaceutically acceptable salt,

for use in a method for preventing, inhibiting or treating a disease ina patient suffering thereof, said disease involving a deregulated p53.

According to a more particular aspect of said embodiment, an additionalsubject-matter of the present invention is a compound of formula (If)

wherein:

R3 is a hydrogen atom or a methyl group,

R13 is a methoxy group and can be in meta or para position,

R14, which can be in meta or para position, represents

-   -   a —CONHR6 group,    -   a triazolyl or imidazolyl group, said groups being optionally        substituted by a chlorine atom, a —(CH₂)_(n)NR7R8 group or a        hydroxy(C₁-C₄)alkyl group,

R6 represents a (C₁-C₆)alkyl group optionally substituted by a hydroxylgroup,

or a —(CH₂)_(n)NR7R8 group,

R7 and R8 independently represent a (C₁-C₄)alkyl group or form togetherwith the nitrogen atom to which they are attached a saturated orinsaturated 5- or 6-membered ring optionally containing a furtherheteroatom chosen among nitrogen and oxygen and optionally substitutedby a methyl group, and said ring being chosen among a imidazolyl, amorpholinyl, a piperidinyl, a pyrrolidinyl or 4-methylpiperazinyl group,

or one of its pharmaceutically acceptable salt,

for use in a method for preventing, inhibiting or treating a disease ina patient suffering thereof, said disease involving a deregulated p53.

According to a preferred embodiment of the present invention, thecompound for use in a method for preventing, inhibiting or treatingdiseases involved with p53 expression, is chosen from one of thecompounds as listed in table I below.

The chemical structures and spectroscopic data of some compounds offormula (I) of the invention are illustrated respectively in thefollowing Table I and Table II.

TABLE I (I)

Compound numbers Formulae (Ia)  1

 2

 3

 4

 5

 6

 7

 8

 9

 10

 11

 12

 13

 14

 15

 16

 17

 18

 19

 20

 21

 22

 23

 24

 25

127

128

129

(Ib)  26

 27

 28

 29

 30

 31

 32

 33

 34

 35

 36

130

131

132

133

134

135

(Ic)  37

 38

 39

 40

 41

 42

 43

 44

136

(Id)  45

 46

 47

 48

 49

 50

 51

 52

 53

 54

 55

 56

 57

 58

 59

 60

(Ie)  61

 62

 63

 64

 65

 66

 67

 68

 69

 70

 71

 72

 73

 74

 75

 76

 77

 78

 79

 80

 81

 82

 83

 84

 85

 86

 87

 88

 89

 90

 91

 92

(If)  93

 94

 95

 96

 97

 98

 99

100

101

102

103

104

105

106

107

108

109

110

111

112

113

114

115

116

117

118

119

120

121

122

123

124

125

126

137

138

139

140

TABLE II Ex Characterizations 1 ¹H NMR (300 MHz, DMSO-d6) δ 8.52 (d, J =5.7 Hz, 2H), 8.39 (s, 1H), 8.35-8.30 (m, 1H), 7.55 (d, J = 6.0 Hz, 2H),7.50 (d, J = 16.7 Hz, 1H), 7.32-7.27 (m, 5H), 7.20 (d, J = 7.6 Hz, 2H),7.15 (d, J = 11.4 Hz, 1H), 7.05 (d, J = 8.2 Hz, 1H), 3.22 (q, J = 6.9Hz, 2H), 1.58 (heptuplet, J = 6.6 Hz, 1H), 1.37 (q, J = 6.9 Hz, 2H),0.87 (d, J = 6.6 Hz, 6H). ¹³C NMR (75 MHz, DMSO-d6) δ 166.3, 150.0,143.6, 143.4, 137.2, 136.1, 133.3, 129.7, 129.1, 125.8, 120.9, 119.0,118.8, 118.2, 117.5, 115.8, 115.6, 38.1, 37.4, 25.3, 22.5. [M + H]⁺ =386.2 2 ¹H NMR (300 MHz, CDCl₃) δ 8.70 (s, 1H), 8.51 (d, J = 4.9 Hz,2H), 7.61 (s, 1H), 7.30-7.14 (m, 8H), 7.08-7.00 (m, 2H), 6.89 (d, J =16.4 Hz, 1H), 6.58 (s, 1H), 3.52-3.48 (m, 2H), 2.59-2.40 (m, 6H),1.69-1.65 (m, 2H), 0.95 (t, J = 7.1 Hz, 6H). ¹³C NMR (75 MHz, CDCl₃) δ167.1, 150.3, 144.7, 143.6, 143.5, 137.6, 136.5, 133.3, 130.0, 129.4,126.3, 121.1, 120.3, 120.2, 119.1, 118.8, 117.1, 116.9, 53.4, 46.9,41.4, 25.0, 11.6. [M + H]⁺ = 429.2 3 ¹H NMR (300 MHz, CDCl₃) δ 8.55 (bs,2H), 7.96 (s, 1H), 7.55 (s, 1H), 7.42-7.03 (m, 10H), 6.96 (d, J = 16.2Hz, 1H), 6.53 (s, 1H), 3.87 (s, 2H), 2.62 (q, J = 7.0 Hz, 4H), 1.10 (t,J = 7.1 Hz, 6H). [M + H]⁺ = 425.2 4 ¹H NMR (300 MHz, CDCl₃) δ 8.52 (bs,2H), 7.73 (s, 1H), 7.50 (s, 1H), 7.38-7.05 (m, 9H), 6.95 (d, J = 16.2Hz, 1H), 6.55 (s, 1H), 2.89 (t, J = 7.4 Hz, 2H), 2.52 (s, 1H), 1.98 (t,J = 6.9 Hz, 2H), 1.23 (t, J = 7.0 Hz, 2H). [M + H]⁺ = 398 5 [M + H]⁺ =386 6 [M + H]⁺ = 443 7 ¹H NMR (300 MHz, CDCl₃) δ 8.47 (d, J = 4.9 Hz,2H), 7.58 (s, 1H), 7.46 (dd, J = 8.4, 2.0 Hz, 1H), 7.36-7.31 (m, 1H),7.27 (d, J = 5.2 Hz, 2H), 7.25-7.03 (m, 5H), 7.00-6.78 (m, 1H), 6.11 (s,1H), 5.71 (s, 1H), 3.38 (q, J = 6.5 Hz, 2H), 2.22 (s, 3H), 1.62(heptuplet, J = 6.7 Hz, 1H), 1.43 (q, J = 6.1 Hz, 2H), 0.83 (d, J = 6.6Hz, 6H) [M + H]⁺ = 400 8 ¹H NMR (300 MHz, DMSO) δ 8.60 (s, 1H),8.50-8.45 (m, 2H), 8.15 (t, J = 6.0 Hz, 1H), 7.75 (d, J = 8.7 Hz, 2H),7.56 (d, J = 4.3 Hz, 2H), 7.50-7.14 (m, 6H), 7.08 (d, J = 8.5 Hz, 2H),3.26 (q, J = 6.4 Hz, 2H), 1.60 (heptuplet, J = 6.6 Hz, 1H), 1.40 (d, J =7.0 Hz, 2H), 0.90 (d, J = 6.6 Hz, 6H). 9 ¹H NMR (300 MHz, C₅D₅N) δ 8.43(s, 1H), 8.23 (d, J = 5.8, 2H), 7.89 (s, 1H), 7.48 (d, J = 7.3, 1H),7.10-6.75 (m, 11H), 6.68 (d, J = 16.4, 1H). ¹³C NMR (75 MHz, C₅D₅N) δ170.4, 151.2, 145.4, 145.3, 138.6, 137.6, 134.3, 130.8, 130.3, 127.0,121.8, 121.0, 120.4, 120.2, 119.0, 117.9, 117.4. [M + H]⁺ = 316 10 [M +H]⁺ = 398 11 [M + H]⁺ = 425 12 [M + H]⁺ = 398 13 ¹H NMR (300 MHz, CDCl₃)δ 8.71 (s, 1H), 8.50 (s, 2H), 7.39 (d, J = 7.7 Hz, 2H), 7.35-7.13 (m,7H), 7.06 (d, J = 8.1 Hz, 2H), 6.81 (d, J = 16.6 Hz, 1H), 6.68 (s, 1H),3.52 (s, 2H), 2.54 (m, 6H), 1.71 (s, 2H), 0.97 (t, J = 6.8 Hz, 6H). [M +H]⁺ = 429.2 14 ¹H NMR (300 MHz, CDCl₃) δ 8.47 (d, J = 5.0 Hz, 2H),7.67-7.45 (m, 4H), 7.42-7.06 (m, 4H), 7.06-6.94 (m, 3H), 6.88 (d, J =17.4 Hz, 1H), 5.72 (s, 1H), 3.55-3.45 (m, 2H), 2.36-2.26 (m, 2H), 2.22(s, 3H), 1.69 (m, 6H), 1.20 (t, 6H). [M + H]⁺ = 443 15 ¹H NMR (300 MHz,CDCl₃) δ 8.48 (d, J = 5.8 Hz, 2H), 7.66-7.31 (m, 6H), 7.30-7.15 (m, 3H),7.06-6.79 (m, 2H), 5.98 (s, 1H), 5.70 (s, 1H), 3.40 (q, J = 6.6 Hz, 2H),2.23 (s, 3H), 1.62 (quint, J = 6.5 Hz, 2H), 1.44 (q, J = 7.1 Hz, 2H),0.89 (d, J = 6.5 Hz, 6H). [M + H]⁺ = 400 16 ¹H NMR (300 MHz, CDCl₃) δ8.50 (s, 2H), 7.52 (s, 1H), 7.38-7.18 (m, 7H), 7.18-7.00 (m, 3H), 6.93(d, J = 16.2 Hz, 1H), 6.66 (s, 1H), 3.76-3.40 (m, 4H), 3.15-2.91 (m,2H), 2.46-2.12 (m, 2H + 4H), 1.69-1.45 (m, 2H). [M + H]⁺ = 479.2 17 ¹HNMR (300 MHz, CDCl₃) δ 8.59 (s, 2H), 7.57 (s, 1H), 7.38 (s, 4H), 7.30(d, J = 5.5 Hz, 4H), 7.15 (dd, J = 27.1, 6.6 Hz, 2H), 7.01 (d, J = 16.9Hz, 1H), 6.29 (s, 1H), 3.17-3.00 (m, 2H), 2.59-2.20 (m, 6H), 1.74-1.52(m, 6H), 1.52-1.35 (m, 2H). [M + H]⁺ = 477.2 18 ¹H NMR (300 MHz, CDCl₃)δ 8.55 (d, J = 5.8 Hz, 2H), 7.58 (s, 1H), 7.39-7.32 (m, 4H), 7.31-7.24(m, 3H), 7.21 (s, 1H), 7.15 (d, J = 7.7 Hz, 1H), 7.10 (d, J = 7.9 Hz,1H), 6.98 (d, J = 16.3 Hz, 1H), 6.64 (d, J = 8.4 Hz, 1H), 3.15-3.00 (m,2H), 2.52-2.35 (m, 2H + 4H), 1.68-1.53 (m, 2H), 0.96 (t, J = 7.1 Hz,6H). [M + H]⁺ = 465.2 19 ¹H NMR (300 MHz, CDCl₃) δ 8.54 (d, J = 4.4 Hz,2H), 7.61 (s, 1H), 7.38-7.16 (m, 7H), 7.10 (d, J = 7.8 Hz, 2H), 7.05 (d,J = 7.8 Hz, 2H), 6.93 (d, J = 16.3 Hz, 1H), 6.30 (s, 1H), 3.53 (q, J =5.0 Hz, 2H), 2.55-2.50-2.46 (m, 2H), 2.45-2.40 (m, 4H), 1.78-1.72 (s,2H), 1.55-1.50 (s, 4H), 1.48-1.35 (m, 2H). ¹³C NMR (75 MHz, CDCl3) δ167.45, 150.34, 144.72, 143.56, 137.58, 136.62, 133.29, 130.03, 129.37,126.34, 121.07, 120.26, 119.38, 118.61, 117.36, 116.65, 59.28, 54.91,41.23, 26.04, 24.45, 24.26. [M + H]⁺ = 441.1 20 ¹H NMR (300 MHz, CDCl₃)δ 8.50 (d, J = 4.2 Hz, 2H), 8.01 (s, 1H), 7.58 (s, 1H), 7.32-7.13 (m,8H), 7.07 (d, J = 7.9 Hz, 1H), 7.03 (d, J = 7.9 Hz, 1H), 6.90 (d, J =16.3 Hz, 1H), 6.54 (s, 1H), 3.70-3.55 (m, 4H), 3.51 (q, J = 4.8 Hz, 2H),2.50-2.40 (m, 6H), 1.84-1.54 (m, 2H). ¹³C NMR (75 MHz, CDCl3) δ 167.56,150.30, 144.69, 143.82, 143.29, 137.59, 136.47, 133.23, 130.04, 129.54,126.38, 121.07, 120.37, 120.13, 118.82, 116.90, 66.97, 58.68, 53.93,40.63, 24.36. [M + H]⁺ = 443.2 21 ¹H NMR (300 MHz, CDCl₃) δ 8.49 (d, J =3.6 Hz, 2H), 8.23 (s, 1H), 7.56 (s, 1H), 7.39-7.11 (m, 7H), 7.14-6.98(m, 3H), 6.90 (d, J = 15.8 Hz, 1H), 6.53 (s, 1H), 3.49 (s, 2H), 3.23 (s,2H), 2.2.5-2.30 (m, 8H), 2.20 (s, 3H), 1.75-1.70 (m, 2H). ¹³C NMR (75MHz, CDCl3) δ 167.47, 150.25, 144.65, 143.65, 143.38, 137.49, 136.43,133.24, 129.95, 129.35, 126.26, 121.02, 120.13, 119.99, 119.17, 118.56,117.07, 116.68, 58.36, 55.12, 53.37, 46.13, 40.93, 24.40. [M + H]⁺ =456.2 22 ¹H NMR (300 MHz, CDCl₃) δ 8.69 (s, 1H), 8.52 (bs, 2H), 7.59 (s,1H), 7.39-7.15 (m, 8H), 7.09 (d, 8.2 Hz, 1H), 7.06 (d, 8.2 Hz, 1H), 6.93(d, J = 15.5 Hz, 1H), 6.38 (s, 1H), 3.54 (s, 2H), 2.70-2.60 (m, 2H),2.55-2.50 (m, 4H), 1.80-1.70 (m, 6H). [M + H]⁺ = 427.2 23 ¹H NMR (300MHz, CDCl₃) δ 8.54 (s, 2H), 7.36-7.30 (d, J = 4.0 Hz, 2H), 7.29-7.16 (m,4H), 7.11 (d, J = 8.8 Hz, 3H), 7.03 (d, J = 7.5 Hz, 2H), 6.98-6.86 (m,2H), 6.59 (s, 1H), 3.90-3.70 (m, 2H), 3.58-3.31 (m, 2H), 2.50-2.21 (m,4H + 3H). [M + H]⁺ = 399.2 24 ¹H NMR (300 MHz, CDCl₃) δ 8.55 (s, 2H),7.58 (s, 1H), 7.43-7.23 (m, 7H), 7.23-7.04 (m, 3H), 6.97 (d, J = 16.3Hz, 1H), 6.67 (s, 1H), 4.51 (br s, 1H), 3.07-2.91 (m, 2H), 2.43-2.28 (m,2H), 2.26-2.03 (m, 4H), 1.53-1.40 (m, 4H), 1.40-1.31 (m, 2H). [M + H]⁺ =463.2 25 ¹H NMR (300 MHz, CDCl₃) δ 8.57 (s, 2H), 7.58 (s, 1H), 7.43-7.23(m, 7H), 7.23-7.05 (m, 3H), 6.98 (d, J = 15.6 Hz, 1H), 6.55 (s, 1H),4.67 (br s, 1H), 3.13-2.93 (m, 2H), 2.61-2.44 (m, 2H), 2.43-2.17 (m,4H), 1.78-1.51 (m, 4H). [M + H]⁺ = 449.2 127 ¹H NMR (300 MHz, MeOD) δ8.43 (s, 2H), 7.67 (s, 1H), 7.53 (d, J = 5.2 Hz, 2H), 7.42 (d, J = 16.5Hz, 1H), 7.38-7.13 (m, 8H), 7.13-7.07 (m, 1H), 7.09 (d, J = 16.5 Hz,1H). [M + H]⁺ = 352.1 128 ¹H NMR (300 MHz, MeOD) δ 8.47 (dd, J = 4.8,1.5 Hz, 2H), 8.18 (s, 1H), 7.66 (dd, J = 6.7, 2.1 Hz, 2H), 7.58 (dd, J =4.8, 1.5 Hz, 2H), 7.47 (d, J = 16.4 Hz, 1H), 7.40 (t, J = 1.6 Hz, 1H),7.32 (t, J = 7.8 Hz, 1H), 7.24 (dd, J = 6.7, 2.1 Hz, 2H), 7.18-7.11 (m,2H), 7.15 (d, J = 16.4 Hz, 1H), 3.65 (q, J = 7.1 Hz, 2H), 2.86 (t, J =7.4 Hz, 2H), 1.95 (q, J = 7.4 Hz, 2H). 129 ¹H NMR (300 MHz, DMSO) δ 9.72(s, 1H), 8.65 (t, J = 5.3 Hz, 1H), 8.54 (br s, 1H), 7.63 (d, J = 7.7 Hz,1H), 7.59-7.52 (m, 2H), 7.52 (d, J = 16.4 Hz, 1H), 7.41 (s, 1H),7.37-7.30 (m, 3H), 7.29-7.20 (m, 1H), 7.24 (d, J = 16.4 Hz, 1H), 7.12(d, J = 8.1 Hz, 1H), 6.91-6.83 (m, 1H), 3.31-3.21 (m, 2H), 2.61-2.39 (m,6H), 1.74-1.59 (m, 2H), 0.95 (t, J = 6.7 Hz, 6H). [M + H]⁺ = 429.3 26 ¹HNMR (300 MHz, CDCl₃) δ 8.51 (d, J = 4.5, 1H), 7.61 (s, 1H), 7.56 (d, J =7.6, 1H), 7.48 (d, J = 16.1, 1H), 7.41 (d, J = 8.7, 2H), 7.30 (d, J =7.8, 1H), 7.21 (s, 1H), 7.17 (d, J = 7.7, 1H), 7.12-7.01 (m, 5H), 6.97(d, J = 7.8, 1H), 6.87 (s, 1H), 4.19 (s, 1H), 3.70 (t, J = 6.2, 2H),2.84 (t, J = 7.4, 2H), 2.00 (quint, J = 6.7, 2H). ¹³C NMR (75 MHz,CDCl₃) δ 155.6, 149.8, 148.2, 144.0, 142.7, 138.1, 136.9, 132.7, 130.3,130.0, 128.4, 122.4, 122.1, 121.0, 119.5, 118.9, 117.7, 117.3, 61.9,32.2, 22.3. [M + H]⁺ = 398 27 ¹H NMR (300 MHz, MeOD) δ 8.49 (s, 1H),7.81 (m, 1H), 7.65-7.45 (m, 4H), 7.35-7.05 (m, 9H), 3.45 (m, 2H), 3.31(m, 4H), 2.06 (m, 2H), 1.28 (m, 6H), 0.89 (m, 2H). [M + H]⁺ = 429 28 ¹HNMR (300 MHz, CDCl₃) δ 8.59 (d, J = 3.9 Hz, 1H), 7.64 (m, 1H), 7.55 (d,J = 16.1 Hz, 1H), 7.48 (s, 1H), 7.44-7.32 (m, 2H), 7.31-6.94 (m, 8H),6.24 (s, 1H), 6.12 (s, 1H), 3.44 (q, J = 6.1 Hz, 2H), 1.66 (heptuplet, J= 6.6 Hz, 1H), 1.48 (q, J = 7.0 Hz, 2H), 0.93 (d, J = 6.6 Hz, 6H). [M +H]⁺ = 386 29 [M + H]⁺ = 398 30 [M + H]⁺ = 429 31 ¹H NMR (300 MHz, CDCl₃)δ 8.58 (d, J = 3.0 Hz, 1H), 7.85-6.73 (m, 12H), 6.25 (t, J = 5.3 Hz,1H), 5.75 (s, 1H), 3.46 (q, J = 6.7 Hz, 2H), 2.27 (s, 3H), 1.68(heptuplet, J = 6.7 Hz, 1H), 1.50 (q, J = 7.1 Hz, 2H), 0.94 (q, J = 6.6Hz, 6H). [M + H]⁺ = 400 32 ¹H NMR (300 MHz, CDCl₃) δ 8.59 (d, J = 5.5Hz, 1H), 7.81-7.48 (m, 6H), 7.47-7.32 (m, 3H), 7.19 (d, J = 8.8 Hz, 2H),7.16-7.02 (m, 2H), 6.98 (s, 1H), 6.09 (s, 1H), 3.77 (t, J = 6.1 Hz, 2H),2.92 (t, J = 7.3 Hz, 2H), 2.04-1.98 (m, 2H). [M + H]⁺ = 398 33 [M + H]⁺= 429 34 ¹H NMR (300 MHz, CDCl₃) δ 8.59 (d, J = 4.2 Hz, 1H), 7.70-7.46(m, 5H), 7.45-7.19 (m, 5H), 7.16-6.99 (m, 3H), 6.01 (s, 1H), 5.93 (s,1H), 3.47 (q, J = 5.9 Hz, 2H), 1.68 (heptuplet, J = 6.6 Hz, 1H), 1.51(q, J = 7.6 Hz, 2H), 0.96 (d, J = 6.6 Hz, 6H). [M + H]⁺ = 386 35 [M +H]⁺ = 429 36 [M + H]⁺ = 386 130 ¹H NMR (300 MHz, MeOD) δ 8.52-8.46 (m,1H), 7.81 (td, J = 7.7, 1.7 Hz, 1H), 7.64 (d, J = 7.9 Hz, 1H), 7.58-7.50(m, 2H), 7.36-7.23 (m, 6H), 7.22-7.15 (m, 2H), 7.19 (d, J = 16.3 Hz,1H), 7.19 (s, 1H), 7.13-7.07 (m, 1H), 3.42 (t, J = 6.7 Hz, 2H),2.91-2.41 (m, 10H), 2.38 (s, 3H), 1.89-1.77 (m, 2H). 131 ¹H NMR (300MHz, MeOD) δ 8.51-8.46 (m, 1H), 7.80 (td, J = 7.8, 1.8 Hz, 1H),7.66-7.58 (m, 2H), 7.53 (d, J = 16.4 Hz, 1H), 7.36-7.24 (m, 6H), 7.18(d, J = 16.4 Hz, 1H), 7.18 (s, 1H), 7.13-7.07 (m, 1H), 3.47 (t, J = 6.6Hz, 2H), 3.15-3.06 (m, 4H), 3.05-2.97 (m, 2H), 2.04-1.92 (m, 6H). 132 ¹HNMR (300 MHz, MeOD) δ 8.50-8.45 (m, 1H), 7.77 (td, J = 7.7, 1.8 Hz, 1H),7.63-7.57 (m, 2H), 7.51 (d, J = 16.3 Hz, 1H), 7.35-7.21 (m, 6H), 7.16(d, J = 16.3 Hz, 1H), 7.16 (s, 1H), 7.12-7.06 (m, 1H), 3.40 (t, J = 6.7Hz, 2H), 2.72-2.55 (m, 6H), 1.95-1.83 (m, 2H), 1.72-1.59 (m, 4H),1.54-1.41 (m, 2H). 133 ¹H NMR (300 MHz, MeOD) δ 8.51-8.47 (m, 1H),7.65-7.59 (m, 1H), 7.59-7.57 (m, 1H), 7.54 (d, J = 16.4 Hz, 1H),7.43-7.24 (m, 6H), 7.23-7.18 (m, 1H), 7.19 (d, J = 16.3 Hz, 1H),7.14-7.08 (m, 1H), 2.95 (t, J = 6.5 Hz, 2H), 2.75-2.61 (m, 6H),1.77-1.62 (m, 2H), 1.07 (t, J = 7.2 Hz, 6H). 134 ¹H NMR (300 MHz, MeOD)δ 8.50-8.44 (m, 1H), 7.76 (td, J = 7.7, 1.7 Hz, 1H), 7.63-7.55 (m, 2H),7.51 (d, J = 16.3 Hz, 1H), 7.36-7.20 (m, 6H), 7.16 (d, J = 16.3 Hz, 1H),7.16 (s, 1H), 7.12-7.06 (m, 1H), 3.65 (t, J = 4.5 Hz, 4H), 3.40 (t, J =6.8 Hz, 2H), 2.55-2.41 (m, 6H), 1.81 (dt, J = 14.0, 6.9 Hz, 2H). 135 ¹HNMR (300 MHz, MeOD) δ 8.51-8.46 (m, 1H), 7.78 (td, J = 7.7, 1.8 Hz, 1H),7.63-7.56 (m, 2H), 7.52 (d, J = 16.3 Hz, 1H), 7.42-7.22 (m, 6H), 7.18(d, J = 16.3 Hz, 1H), 7.18 (s, 1H), 7.11 (dd, J = 7.9, 1.1 Hz, 1H), 2.93(t, J = 6.6 Hz, 2H), 2.61-2.45 (m, 6H), 1.79-1.67 (m, 2H), 1.66-1.55 (m,4H), 1.51-1.38 (m, 2H). 37 ¹H NMR (300 MHz, CDCl₃) δ 8.62 (s, 1H), 7.59(s, 1H), 7.49 (d, J = 7.3 Hz, 2H), 7.24 (m, Hz, 6H), 7.06 (m, 5H), 6.10(s, 1H), 3.55 (q, 2H), 2.74-2.40 (m, 6H), 1.75 (m, 2H), 1.01 (t, 6H).[M + H]⁺ = 512 38 [M + H]⁺ = 539 39 ¹H NMR (300 MHz, CDCl₃) δ 8.22 (s,1H), 7.58 (s, 1H), 7.44-7.17 (m, 9H), 7.11 (d, J = 7.3 Hz, 2H), 7.03 (d,J = 2.6 Hz, 2H), 6.16 (s, 1H), 3.55 (q, J = 5.0 Hz, 3H), 2.60-2.40 (m,10H), 2.25 (s, 3H), 1.88-1.55 (m, 2H). [M + H]⁺ = 539 40 ¹H NMR (300MHz, CDCl₃) δ 8.59 (s, 1H), 7.52 (s, 1H), 7.43 (d, J = 8.7 Hz, 2H), 7.27(d, J = 7.0 Hz, 2H), 7.23 (d, J = 7.3 Hz, 2H), 7.18 (s, 1H), 7.13-6.92(m, 4H), 6.89 (d, J = 8.7 Hz, 2H), 3.82 (s, 3H), 3.59-3.47 (m, 2H),2.62-2.46 (m, 6H), 1.72 (quint, J = 5.7 Hz, 2H), 0.99 (t, J = 7.2 Hz,6H). ¹³C NMR (75 MHz, CDCl₃) δ 167.2, 159.5, 144.0, 143.2, 139.1, 136.4,130.2, 129.8, 129.4, 128.6, 127.9, 119.8, 118.9, 117.6, 116.6, 114.3,55.5, 53.3, 46.9, 41.3, 25.1, 11.6. [M + H]⁺ = 458.3 41 ¹H NMR (300 MHz,CDCl₃) δ 7.45-7.31 (m, 4H), 7.25-7.20 (m, 1H), 7.19 (s, 1H), 7.17-7.10(m, 3H), 7.09-6.98 (m, 1H), 6.96-6.75 (m, 4H), 3.75 (s, 3H), 1.59(heptuplet, J = 13.3, 6.6 Hz, 1H), 1.42 (q, J = 8.7, 7.1 Hz, 2H), 0.87(d, J = 6.6 Hz, 6H) ¹³C NMR (75 MHz, CDCl₃) δ 167.6, 159.5, 144.0,142.8, 139.2, 136.4, 130.1, 129.8, 129.6, 129.1, 128.7, 128.5, 127.9,126.5, 120.0, 119.8, 118.6, 117.7, 116.5, 116.3, 114.3, 55.4, 38.6,38.5, 26.1, 22.6 [M + H]⁺ = 415 42 [M + H]⁺ = 415 43 [M + H]⁺ = 429 44¹H NMR (300 MHz, CDCl₃) δ 10.77 (s, 1H), 8.15 (s, 1H), 7.75 (s, 1H),7.66 (d, J = 9.0 Hz, 1H), 7.39 (d, J = 8.3 Hz, 2H), 7.30-7.05 (m, 3H),7.0-6.90 (m, 3H), 6.86 (s, 1H), 6.84 (d, J = 8.5 Hz, 2H), 3.77 (s, 3H),3.55-3.49 (m, 2H), 2.95-2.80 (m, 6H), 2.26 (s, 3H), 2.15-2.05 (m, 2H),1.30 (t, J = 7.2 Hz, 6H). [M + H]⁺ = 472 136 ¹H NMR (300 MHz, MeOD) δ7.57-7.53 (m, 2H), 7.52 (s, 1H), 7.36-7.21 (m, 8H), 7.15-7.10 (m, 3H),7.04 (ddd, J = 7.9, 2.2, 1.0 Hz, 1H), 3.39 (t, J = 6.7 Hz, 2H),2.68-2.58 (m, 6H), 1.87-1.74 (m, 2H), 1.05 (t, J = 7.2 Hz, 6H). 45 ¹HNMR (300 MHz, CDCl₃) δ 9.55 (s, 1H), 7.48 (d, J = 7.7 Hz, 1H), 7.30-7.21(m, 3H), 7.15 (d, J = 9.1 Hz, 2H), 7.10 (d, J = 9.1 Hz, 2H), 6.78 (t, J= 7.3 Hz, 1H), 3.44 (t, J = 5.9 Hz, 2H), 2.62 (t, J = 5.9 Hz, 2H), 2.54(q, J = 7.1 Hz, 4H), 1.02 (t, J = 7.1 Hz, 6H). ¹³C NMR (75 MHz, CDCl₃) δ169.5, 145.2, 143.9, 140.8, 132.2, 127.8, 122.3, 121.4, 119.1, 118.7,115.6, 51.3, 46.9, 37.2, 12.1. [M + H]⁺ = 396 46 ¹H NMR (300 MHz, CDCl₃)δ 9.83 (s, 1H), 8.84 (s, 1H), 7.42 (d, J = 7.3 Hz, 1H), 7.28 (t, J = 7.1Hz, 2H), 7.20-7.07 (m, 5H), 6.75 (t, J = 6.7 Hz, 1H), 3.52 (q, J = 5.2Hz, 2H), 2.82-2.40 (m, 6H), 1.80-1.70 (m, 2H), 1.03 (t, J = 7.1 Hz, 6H)¹³C NMR (75 MHz, CDCl₃) δ 169.4, 145.3, 143.7, 140.9, 132.0, 127.9,122.2, 121.3, 119.0, 118.7, 118.2, 115.4, 53.6, 46.9, 41.2, 24.8, 11.5[M + H]⁺ = 410 47 ¹H NMR (300 MHz, CDCl₃) δ 9.50 (s, 1H), 7.47 (d, J =7.4 Hz, 1H), 7.35-7.23 (m, 2H), 7.18 (d, J = 9.0 Hz, 2H), 7.14 (d, J =9.0 Hz, 2H), 6.94 (bs, 1H), 6.80 (t, J = 6.3 Hz, 1H), 3.48 (bs, 2H),2.54 (bs, 2H), 2.28 (s, 6H). [M + H]⁺ = 368 48 ¹H NMR (300 MHz, CDCl₃) δ8.31 (d, J = 8.9, 1H), 7.95 (d, J = 2.8, 1H), 7.61 (t, J = 9.1, 2H),6.97 (d, J = 8.9, 1H), 6.84 (d, J = 8.8, 1H), 6.59 (d, J = 8.5, 1H),4.25 (t, J = 5.4, 2H), 3.92 (s, 3H), 3.79-3.72 (m, 1H), 2.92 (t, J =5.5, 2H), 2.68-2.60 (m, 5H) [M + H]⁺ = 367.2 49 ¹H NMR (300 MHz, CDCl₃)δ 9.90 (s, 1H), 8.74 (s, 1H), 7.58 (d, J = 6.4 Hz, 1H), 7.45-7.21 (m,3H), 7.09 (d, J = 11.1 Hz, 2H), 6.83 (dd, J = 16.4, 7.7 Hz, 2H),3.76-3.40 (m, 2H), 3.13-2.41 (m, 2H + 4H), 2.10-1.69 (m, 2H), 1.15 (t, J= 7.2 Hz, 6H). [M + H]⁺ = 410.2 50 ¹H NMR (300 MHz, CDCl₃) δ 9.64 (s,1H), 8.74 (s, 1H), 7.49-7.30 (m, 6H), 7.23-7.08 (m, 4H), 6.95 (d, J =8.9 Hz, 2H), 6.66 (t, J = 7.4 Hz, 1H), 5.06 (s, 2H), 3.61-3.45 (m, 2H),2.73-2.47 (m, 2H + 4H), 1.84-1.71 (m, 2H), 1.07 (t, J = 7.1 Hz, 6H). ¹³CNMR (75 MHz, CDCl₃) δ 169.7, 155.1, 147.6, 137.3, 135.0, 132.0, 128.7,128.1, 127.8, 127.7, 124.4, 116.7, 116.5, 115.7, 114.3, 70.5, 53.5,46.8, 41.0, 24.8, 11.5. [M + H]⁺ = 432.2 51 ¹H NMR (300 MHz, CDCl₃) δ9.77 (s, 1H), 8.77 (s, 1H), 7.43 (d, J = 7.8 Hz, 1H), 7.35 (t, J = 8.7Hz, 1H), 7.30-7.14 (m, 5H), 6.99 (t, J = 7.1 Hz, 1H), 6.73 (t, J = 7.4Hz, 1H), 3.60-3.46 (m, 2H), 2.68-2.50 (m, 2H + 4H), 1.84-1.69 (m, 2H),1.05 (t, J = 7.1 Hz, 6H). ¹³C NMR (75 MHz, CDCl₃) δ 169.5, 145.7, 141.9,131.9, 129.3, 127.8, 122.2, 120.8, 118.3, 117.6, 115.3, 53.5, 46.8,41.0, 24.8, 11.4. [M + H]⁺ = 326.3 52 ¹H NMR (300 MHz, CDCl₃) δ 9.50 (s,1H), 8.75 (s, 1H), 7.41 (d, J = 7.8 Hz, 1H), 7.25-7.14 (m, 1H), 7.06 (d,J = 8.3 Hz, 2H), 7.00-6.89 (m, 1H), 6.80 (d, J = 8.2 Hz, 2H), 6.66 (t, J= 7.4 Hz, 1H), 3.69-3.42 (m, 2H), 2.71-2.52 (m, 2H + 4H), 1.89-1.72 (m,2H), 1.07 (t, J = 7.1 Hz, 6H). [M + H]⁺ = 342.3 53 [M + H]⁺ = 414 54[M + H]⁺ = 351 55 [M + H]⁺ = 405 56 [M + H]⁺ = 364 57 [M + H]⁺ = 397 58[M + H]⁺ = 342 59 [M + H]⁺ = 356 60 ¹H NMR (300 MHz, CDCl₃) δ 9.23 (s,1H), 7.40 (s, 1H), 7.31 (d, J = 8.3 Hz, 1H), 7.20-7.00 (m, 3H), 6.97 (t,J = 8.1 Hz, 1H), 6.76-6.61 (m, 3H), 6.47 (d, J = 7.5 Hz, 1H), 6.22 (t, J= 8.7 Hz, 1H), 6.16 (s, 1H), 4.01 (m, 2H), 3.70 (s, 3H), 3.32 (m, 2H),2.08 (m, 2H). [M + H]⁺ = 351 61 [M + H]⁺ = 405 62 [M + H]⁺ = 314 63 [M +H]⁺ = 368 64 [M + H]⁺ = 328 65 ¹H NMR (300 MHz, CDCl₃) δ 7.62 (s, 1H),7.50 (dd, J = 1.9, 8.5 Hz, 1H), 7.23-7.14 (m, 2H), 6.69-6.59 (m, 2H),6.54 (dd, J = 2.2, 8.1 Hz, 1H), 6.30 (d, J = 5.4 Hz, 1H), 5.73 (s, 1H),3.75 (s, 3H), 3.40 (q, J = 7.0 Hz, 2H), 2.23 (s, 3H), 1.69 (heptuplet, J= 6.6 Hz, 1H), 1.50 (q, J = 7.0 Hz, 2H), 0.93 (d, J = 6.6 Hz, 6H). ¹³CNMR (75 MHz, CDCl₃) 167.4, 160.9, 144.7, 143.6, 130.4, 130.1, 126.7,125.8, 125.7, 115.5, 112.2, 107.7, 105.4, 55.4, 38.8, 38.5, 26.2, 22.7,18.0. [M + H]⁺ = 327 66 ¹H NMR (300 MHz, CDCl₃) δ 7.57 (d, J = 1.5 Hz,1H), 7.52-7.32 (m, 2H), 7.07 (d, J = 8.9 Hz, 2H), 6.87 (d, J = 8.9 Hz,2H), 5.96 (s, 1H), 5.46 (s, 1H), 3.79 (s, 3H), 3.42 (q, J = 7.4 Hz, 2H),2.25 (s, 3H), 1.65 (quint, J = 7.4 Hz, 1H), 1.46 (q, J = 7.4 Hz, 2H),0.92 (d, J = 6.6 Hz, 6H). [M + H]⁺ = 327 67 ¹H NMR (300 MHz, CDCl₃) δ7.65 (s, 1H), 7.51 (d, J = 8.4 Hz, 1H), 7.16 (d, J = 8.6 Hz, 3H), 7.07(d, J = 8.9 Hz, 2H), 6.02 (s, 1H), 5.65 (s, 1H), 3.47 (q, J = 7.0 Hz,2H), 2.29 (s, 3H), 1.75-1.64 (m, 1H), 1.51 (q, J = 7.0 Hz, 2H), 0.96 (d,J = 6.6 Hz, 6H). [M + H]⁺ = 381 68 [M + H]⁺ = 356 69 ¹H NMR (300 MHz,CDCl₃) δ 8.48 (s, 1H), 7.68 (s, 1H), 7.56 (d, J = 8.4 Hz, 1H), 7.26-7.17(m, 2H), 6.74-6.60 (m, 2H), 6.57 (d, J = 8.4 Hz, 1H), 5.59 (s, 1H), 3.79(s, 3H), 3.56 (q, J = 5.3 Hz, 2H), 2.71-2.51 (m, 2H + 4H), 2.29 (s, 3H),1.78 (quint, J = 5.9 Hz, 2H), 1.07 (t, J = 7.1 Hz, 6H). ¹³C NMR (75 MHz,CDCl₃) δ 167.0, 160.8, 144.4, 143.7, 130.3, 130.1, 126.8, 125.9, 125.6,115.4, 112.1, 107.6, 105.3, 58.6, 55.4, 46.9, 27.0, 18.6, 18.0, 11.5.[M + H]⁺ = 370 70 ¹H NMR (300 MHz, CDCl₃) δ 8.34 (s, 1H), 7.62 (s, 1H),7.47 (d, J = 8.8 Hz, 1H), 7.08 (d, J = 8.8 Hz, 2H), 6.88 (d, J = 8.7 Hz,3H), 5.44 (s, 1H), 3.80 (s, 3H), 3.52 (q, J = 5.3 Hz, 2H), 2.61 (t, J =5.1 Hz, 2H), 2.57 (q, J = 7.1 Hz, 4H), 2.26 (s, 3H), 1.73 (quint, J =5.7 Hz, 2H), 1.04 (t, J = 7.1 Hz, 6H). [M + H]⁺ = 370 71 ¹H NMR (300MHz, CDCl₃) δ 8.55 (s, 1H), 7.68 (s, 1H), 7.56 (d, J = 8.5 Hz, 1H),7.18-7.12 (m, 3H), 7.06 (d, J = 9.0 Hz, 2H), 5.81 (s, 1H), 3.54 (qd, J =5.5 Hz, 2H), 2.70-2.60 (m, 2H), 2.62 (q, J = 7.1 Hz, 4H), 2.27 (s, 3H),1.77 (heptuplet, J = 5.6 Hz, 2H), 1.07 (t, J = 7.1 Hz, 6H). [M + H]⁺ =424 72 [M + H]⁺ = 382 73 [M + H]⁺ = 325 74 [M + H]⁺ = 325 75 ¹H NMR (300MHz, CDCl₃) δ 8.05 (s, 1H), 7.61 (d, J = 8.6 Hz, 2H), 7.10 (d, J = 8.8Hz, 2H), 6.85 (dd, J = 12.4, 8.8 Hz, 4H), 6.08 (s, 1H), 3.79 (s, 3H),3.49 (dd, J = 11.4, 5.7 Hz, 3H), 2.43 (t, J = 6.2 Hz, 3H), 2.24 (s, 6H),1.73 (dd, J = 12.1, 6.0 Hz, 3H), 1.25 (s, 6H). 76 ¹H NMR (300 MHz,CDCl₃) δ 10.20 (s, 1H), 9.91 (s, 1H), 7.60 (s, 1H), 7.40 (d, J = 8.4 Hz,1H), 7.03 (d, J = 8.4 Hz, 2H), 6.95-6.80 (m, 3H), 5.34 (s, 1H), 3.80 (s,3H), 2.17 (s, 3H) 77 ¹H NMR (300 MHz, CDCl₃) δ 7.59 (d, J = 8.7 Hz, 2H),7.06 (d, J = 8.9 Hz, 1H), 6.84 (d, J = 8.9 Hz, 2H), 6.79 (d, J = 8.7 Hz,2H), 3.41 (q, J = 5.3 Hz, 2H), 2.58 (t, J = 7.1 Hz,, 2H), 2.51 (q, J =7.1 Hz, 4H), 0.98 (t, J = 7.1 Hz, 6H). [M + H]⁺ = 342 78 [M + H]⁺ = 35279 [M + H]⁺ = 352 80 ¹H NMR (300 MHz, CDCl₃) δ 7.84 (s, 1H), 7.53 (d, J= 8.7 Hz, 2H), 7.09 (d, J = 8.0 Hz, 2H), 7.06 (d, J = 8.0 Hz, 2H), 7.03(d, J = 8.0 Hz, 2H), 5.96 (s, 1H), 3.82 (s, 2H), 2.57 (q, J = 7.1 Hz,4H), 1.06 (t, J = 7.1 Hz, 6H). [M + H]⁺ = 406 81 M + H]⁺ = 313 82 ¹H NMR(300 MHz, CDCl₃) δ 7.59 (d, J = 8.7 Hz, 2H), 7.09 (d, J = 8.9 Hz, 2H),6.86 (d, J = 8.9 Hz, 2H), 6.81 (d, J = 8.7 Hz, 2H), 5.80 (s, 1H), 3.79(s, 3H), 3.42 (q, J = 7.1 Hz, 2H), 1.65 (quint, J = 6.6 Hz, 1H), 1.47(q, 7.1 Hz, 1H), 0.92 (d, J = 6.6 Hz, 6H). M + H]⁺ = 313 83 ¹H NMR (300MHz, CDCl₃) δ 7.72 (d, J = 8.6 Hz, 2H), 7.40 (d, J = 7.8 Hz, 1H), 7.27(d, J = 7.8 Hz, 1H), 7.20 (t, J = 7.8 Hz, 1H), 7.08 (d, J = 8.6 Hz, 2H),6.96 (t, J = 7.8 Hz, 1H), 6.24 (s, 1H), 6.13 (s, 1H), 3.52-3.39 (q, J =6.6 Hz, 2H), 1.67 (hept, J = 6.6 Hz, 1H), 1.50 (q, J = 6.6 Hz, 2H), 0.94(d, J = 6.6 Hz, 6H). [M + H]⁺ = 367.1 84 ¹H NMR (300 MHz, CDCl₃) δ 7.69(d, J = 8.7 Hz, 1H), 7.27 (d, J = 7.5 Hz, 1H), 7.06 (d, J = 8.7 Hz, 1H),7.04-6.94 (m, 2H), 6.82 (d, J = 7.5 Hz, 1H), 6.27 (s, 1H), 6.07 (s, 1H),3.46 (q, J = 6.6 Hz, 2H), 1.68 (hept, J = 6.6 Hz, 1H), 1.50 (q, J = 6.6Hz, 2H), 0.95 (d, J = 6.6 Hz, 1H). ¹³C NMR (75 MHz, CDCl₃) δ 167.1,150.3, 145.4, 143.5, 130.6, 128.7, 127.3, 116.8, 116.6, 114.0, 111.0,38.7, 38.5, 26.1, 22.6. [M + H]⁺ = 367.1 85 ¹H NMR (300 MHz, CDCl₃) δ7.69 (d, J = 8.6 Hz, 2H), 7.21-7.07 (m, 2H), 7.02 (d, J = 8.6 Hz, 2H),6.08-5.90 (m, 2H), 3.48 (q, J = 6.6 Hz, 2H), 1.70 (hept, J = 6.6 Hz,1H), 1.51 (q, J = 6.6 Hz, 2H), 0.96 (d, J = 6.6 Hz, 6H). [M + H]⁺ =367.1 86 ¹H NMR (300 MHz, CDCl₃) δ 7.66 (s, 1H), 7.54 (d, J = 8.3 Hz,1H), 7.45-7.37 (m, 1H), 7.31-7.16 (m, 4H), 6.97-6.89 (m, 1H), 6.08 (s,1H), 3.47 (q, J = 6.6 Hz, 2H), 2.29 (s, 3H), 1.68 (hept, J = 6.6 Hz,1H), 1.51 (q, J = 6.6 Hz, 2H), 0.96 (d, J = 6.6 Hz, 6H). ¹³C NMR (75MHz, CDCl₃) δ 167.2, 143.5, 143.1, 135.8, 130.1, 129.1, 128.5, 128.4,128.1, 127.6, 125.8, 122.1, 121.3, 118.4, 117.5, 38.8, 38.5, 26.1, 22.6,17.8. [M + H]⁺ = 381.1 87 ¹H NMR (300 MHz, CDCl₃) δ 7.66 (s, 1H), 7.52(d, J = 8.4 Hz, 1H), 7.47-7.38 (m, 1H), 7.30 (d, J = 8.4 Hz, 1H), 7.24(d, J = 8.4 Hz, 1H), 6.95 (d, J = 8.0 Hz, 1H), 6.89 (s, 1H), 6.82 (d, J= 8.0 Hz, 1H), 6.07 (s, 1H), 5.75 (s, 1H), 3.47 (q, J = 6.6 Hz, 2H),2.29 (s, 3H), 1.69 (hept, J = 6.6 Hz, 1H), 1.51 (q, J = 6.6 Hz, 2H),0.96 (d, J = 6.6 Hz, 6H). ¹³C NMR (75 MHz, CDCl₃) δ 167.2, 150.3, 144.3,143.6, 130.6, 130.3, 129.1, 128.5, 128.0, 127.1, 125.7, 116.7, 116.6,113.6, 110.9, 38.8, 38.5, 26.1, 22.6, 18.0. [M + H]⁺ = 381.1 88 ¹H NMR(300 MHz, CDCl₃) δ 8.21 (s, 1H), 7.89 (s, 1H), 7.80 (d, J = 8.3 Hz, 1H),7.37-7.20 (m, 2H), 6.98 (d, J = 8.1 Hz, 1H), 6.90 (s, 1H), 6.80 (d, J =8.0 Hz, 1H), 5.90 (s, 1H), 3.70-3.52 (m, 2H), 3.34-3.00 (m, 2H + 4H),2.29 (s, 3H), 2.25-2.08 (m, 2H), 1.36 (t, J = 7.2 Hz, 6H). ¹³C NMR (75MHz, CDCl₃) δ 167.9, 150.3, 144.2, 143.9, 130.7, 130.5, 126.9, 126.5,116.8, 116.4, 113.5, 111.0, 49.5, 46.2, 41.1, 36.6, 23.9, 18.0, 8.4.[M + H]⁺ = 424.1 89 ¹H NMR (300 MHz, CDCl₃) δ 8.50 (s, 1H), 7.68 (s,1H), 7.54 (d, J = 8.4 Hz, 1H), 7.32 (t, J = 7.1 Hz, 2H), 7.20 (d, J =8.5 Hz, 1H), 7.11 (d, J = 7.5 Hz, 2H), 7.02 (t, J = 7.1 Hz, 1H), 5.61(s, 1H), 3.70-3.40 (m, 2H), 2.76-2.46 (m, 2H + 4H), 2.29 (s, 3H),1.85-1.66 (m, 2H), 1.06 (t, J = 6.8 Hz, 6H). ¹³C NMR (75 MHz, CDCl₃) δ167.0, 144.7, 142.2, 130.1, 129.6, 126.5, 125.9, 125.1, 122.3, 119.9,114.5, 53.3 (CH2), 46.9 (CH2 Et), 25.1 (CH2), 18.0, 11.5. [M + H]⁺ =339.9 90 ¹H NMR (300 MHz, CDCl₃) δ 7.65 (d, J = 8.4 Hz, 2H), 7.60 (d, J= 6.2 Hz, 1H), 7.22 (d, J = 8.1 Hz, 1H), 7.04 (d, J = 8.3 Hz, 2H),6.82-6.67 (m, 2H), 6.60 (d, J = 8.1 Hz, 1H), 6.18 (s, 1H), 6.03 (s, 1H),4.07 (t, J = 6.7 Hz, 2H), 3.80 (s, 3H), 3.53-3.43 (m, 2H), 2.29-2.03 (m,2H). [M + H]⁺ = 351.1 91 ¹H NMR (300 MHz, CDCl₃) δ 7.63 (s, 1H), 7.49(d, J = 8.5 Hz, 1H), 7.26 (d, J = 7.8 Hz, 1H), 7.21 (d, J = 7.8 Hz, 1H),7.09 (s, 1H), 7.01 (s, 1H), 6.75-6.64 (m, 2H), 6.59 (d, J = 8.2 Hz, 1H),6.39 (s, 1H), 5.66 (s, 1H), 4.06 (t, J = 6.8 Hz, 2H), 3.79 (s, 3H), 3.47(q, J = 6.4 Hz, 2H), 2.29 (s, 3H), 2.18-2.05 (quint, J = 6.6 Hz, 2H).¹³C NMR (75 MHz, CDCl₃) δ 160.8, 145.1, 143.2, 130.4, 130.1, 125.9,125.4, 115.0, 112.4, 107.9, 105.8, 55.4, 45.0, 37.3, 31.5, 17.9. [M +H]⁺ = 365.1 92 ¹H NMR (300 MHz, CDCl₃) δ 7.69 (s, 1H), 7.56 (d, J = 8.4Hz, 1H), 7.26 (d, J = 7.8 Hz, 1H), 7.21 (d, J = 7.8 Hz, 1H), 6.81 (br s,1H), 6.73-6.63 (m, 2H), 6.58 (d, J = 8.2 Hz, 1H), 5.60 (s, 1H), 3.80 (s,3H), 3.55 (q, J = 5.7 Hz, 2H), 2.58 (t, J = 5.7 Hz, 2H), 2.33 (s, 6H),2.31 (s, 3H). [M + H]⁺ = 328.3 93 [M + H]⁺ = 475 94 [M + H]⁺ = 489 95[M + H]⁺ = 444 96 [M + H]⁺ = 471 97 [M + H]⁺ = 471 98 [M + H]⁺ = 444 99¹H NMR (300 MHz, CDCl₃) δ 8.05-7.95 (m, 2H), 7.86-7.65 (m, 2H),7.70-7.54 (m, 2H), 7.48-7.30 (m, 5H), 7.15-6.95 (m, 3H), 6.87 (d, J =8.4 Hz, 1H), 3.85 (s, 3H), 3.72 (t, J = 6.5 Hz, 2H), 2.90 (t, J = 6.5Hz, 2H), 1.97 (quint, J = 6.5 Hz, 2H). [M + H]⁺ = 444 100 [M + H]⁺ = 432101 ¹H NMR (300 MHz, CDCl₃) δ 8.41 (s, 1H), 7.88 (s, 1H), 7.76 (d, J =8.7 Hz, 2H), 7.56 (d, J = 14.7 Hz, 2H), 7.36-7.29 (m, 1H), 7.25-6.93 (m,5H), 6.87 (d, J = 8.8 Hz, 2H), 6.71 (d, J = 8.2 Hz, 1H), 5.88 (s, 1H),3.77 (s, 3H), 3.43 (q, J = 6.0 Hz, 2H), 2.55-2.45 (m, 6H), 0.91 (t, J =7.1 Hz, 6H) [M + H]⁺ = 475 102 [M + H]⁺ = 446 103 [M + H]⁺ = 471 104[M + H]⁺ = 444 105 [M + H]⁺ = 475 106 [M + H]⁺ = 489 107 [M + H]⁺ = 446108 [M + H]⁺ = 475 109 [M + H]⁺ = 432 110 [M + H]⁺ = 432 111 [M + H]⁺ =471 112 [M + H]⁺ = 444 113 [M + H]⁺ = 432 114 [M + H]⁺ = 475 115 ¹H NMR(300 MHz, CDCl₃) δ 7.85 (d, J = 8.8 Hz, 2H), 7.53 (d, J = 8.7 Hz, 2H),7.44-7.36 (m, 2H), 7.30-7.11 (m, 2H), 7.08 (d, J = 8.8 Hz, 2H), 6.96 (d,J = 8.9 Hz, 2H), 6.09 (s, 1H), 5.83 (s, 1H), 3.87 (s, 3H), 3.45 (q, J =7.1 Hz, 2H), 1.80-1.63 (m, 1H), 1.49 (q, J = 7.1 Hz, 2H), 0.94 (d, J =6.6 Hz, 6H). [M + H]⁺ = 432 116 ¹H NMR (300 MHz, CDCl₃) δ 7.98 (s, 1H),7.96 (s, 1H), 7.86 (d, J = 8.8 Hz, 2H), 7.57 (d, J = 8.7 Hz, 2H), 7.39(s, 1H), 7.30 (t, J = 8.0 Hz, 1H), 7.13 (d, J = 8.7 Hz, 3H), 7.01 (d, J= 8.0 Hz, 1H), 6.95 (d, J = 8.8 Hz, 2H), 6.13 (s, 1H), 3.91 (s, 2H),3.86 (s, 3H), 2.66 (q, J = 7.1 Hz, 2H), 1.14 (t, J = 7.2 Hz, 6H). [M +H]⁺ = 471 117 [M + H]⁺ = 475 118 [M + H]⁺ = 489 119 [M + H]⁺ = 432 120[M + H]⁺ = 462 121 [M + H]⁺ = 433 122 [M + H]⁺ = 462 123 ¹H NMR (300MHz, CDCl₃) δ 8.62 (s, 1H), 7.94 (d, J = 9.0 Hz, 3H), 7.54 (d, J = 8.6Hz, 2H), 7.40 (s, 1H), 7.23 (d, J = 8.2 Hz, 2H), 7.11-6.95 (m, 4H), 6.43(s, 1H), 3.83 (s, 2H), 2.59 (q, J = 7.1 Hz, 4H), 1.09 (t, J = 7.1 Hz,6H). [M + H]⁺ = 525.1 124 ¹H NMR (300 MHz, CDCl₃) δ 9.74 (s, 1H), 8.23(d, J = 7.8 Hz, 1H), 7.90 (s, 1H), 7.55 (d, J = 8.7 Hz, 2H), 7.47-7.36(m, 2H), 7.24 (t, J = 8.0 Hz, 1H), 7.10 (d, J = 8.8 Hz, 2H), 7.09-6.94(m, 4H), 6.63 (s, 1H), 3.99 (s, 3H), 3.82 (s, 2H), 2.56 (q, J = 7.1 Hz,4H), 1.07 (t, J = 7.1 Hz, 6H). ¹³C NMR (75 MHz, CDCl₃) δ 163.3, 157.4,145.9, 145.6, 138.3, 138.1, 133.5, 133.4, 132.6, 130.6, 122.2, 121.8,121.1, 121.0, 116.0, 111.7, 111.4, 107.9, 56.4, 47.8, 46.9, 11.9. [M +H]⁺ = 471.1 125 ¹H NMR (300 MHz, CDCl₃) δ 8.27 (s, 1H), 8.01 (s, 1H),7.91 (d, J = 7.2 Hz, 2H), 7.61 (d, J = 8.4 Hz, 2H), 7.57-7.45 (m, 3H),7.39 (s, 1H), 7.34 (t, J = 8.2 Hz, 1H), 7.17 (d, J = 8.4 Hz, 3H), 7.05(d, J = 8.4 Hz, 1H), 6.14 (s, 1H), 4.08 (s, 2H), 2.85 (q, J = 7.0 Hz,4H), 1.25 (t, J = 7.0 Hz, 6H). [M + H]⁺ = 441.1 126 ¹H NMR (300 MHz,CDCl₃) δ 8.34 (s, 1H), 7.93 (s, 1H), 7.86 (d, J = 8.9 Hz, 2H), 7.69 (d,J = 8.0 Hz, 1H), 7.61 (d, J = 8.1 Hz, 1H), 7.45 (d, J = 8.5 Hz, 4H),7.39 (s, 1H), 7.21 (s, 1H), 7.00 (d, J = 8.7 Hz, 3H), 6.93 (d, J = 8.8Hz, 3H), 6.13 (br s, 1H), 4.06-3.94 (m, 2H), 3.85 (s, 3H), 3.47-3.34 (m,2H), 2.14-2.01 (m, 2H). [M + H]⁺ = 470.2 137 ¹H NMR (300 MHz, MeOD) δ8.68 (s, 1H), 7.60 (d, J = 8.9 Hz, 2H), 7.52-7.45 (m, 3H), 7.39 (t, J =7.9 Hz, 1H), 7.34 (t, J = 8.1 Hz, 1H), 7.20 (d, J = 2.9 Hz, 1H), 7.17(d, J = 9.0 Hz, 2H), 7.11 (dd, J = 8.2, 3.3 Hz, 2H), 4.22 (s, 2H), 3.85(s, 3H), 2.94 (q, J = 7.2 Hz, 4H), 1.28 (t, J = 7.2 Hz, 6H). 138 ¹H NMR(300 MHz, DMSO) δ 10.13 (s, 1H), 8.42 (s, 1H), 8.38 (s, 1H), 7.68 (d, J= 11.4 Hz, 4H), 7.55-7.46 (m, 2H), 7.43 (t, 1H), 7.13 (d, J = 8.8 Hz,4H), 3.83 (s, 3H), 1.51 (s, 6H). 139 ¹H NMR (300 MHz, MeOD) δ 8.17 (t, J= 5.6 Hz, 1H), 7.69 (d, J = 8.8 Hz, 2H), 7.59 (d, J = 8.8 Hz, 2H),7.52-7.44 (m, 2H), 7.37 (t, J = 7.9 Hz, 1H), 7.14 (d, J = 8.8 Hz, 2H),7.09 (dd, J = 7.8, 2.9 Hz, 1H), 7.02 (d, J = 8.8 Hz, 2H), 3.82 (s, 3H),3.55 (t, J = 6.4 Hz, 2H), 3.40-3.29 (m, 2H), 1.67-1.51 (m, 4H),1.47-1.37 (m, 2H). 140 ¹H NMR (300 MHz, MeOD) δ 8.08 (s, 1H), 7.62 (d, J= 8.7 Hz, 2H), 7.55 (d, J = 8.9 Hz, 2H), 7.48 (d, J = 7.5 Hz, 2H), 7.37(t, J = 7.9 Hz, 1H), 7.13-7.10 (d, J = 8.7 Hz, 2H), 7.10-7.06 (d, J =8.9 Hz, 3H), 4.48 (t, J = 7.0 Hz, 2H), 3.82 (s, 3H), 3.59 (m, 2H),2.21-2.04 (m, 2H).

Among said compounds, compounds (1), (2), (3), (4), (8), (26), (40),(45), (46) and (61) are of particular interest.

The present invention therefore extends to compounds (1), (2), (3), (4),(8), (26), (40), (45), (46) and (61) or one of its pharmaceuticallyacceptable salts for use in a method for preventing, inhibiting ortreating a disease in a patient exhibiting a deregulated p53.

Some of the disclosed compounds are new and form part of the presentinvention: (9), (16), (17), (18), (19), (20), (21), (22), (23), (24),(25), (37), (38), (39), (48), (49), (50), (52), (83), (84), (85), (86),(87), (88), (89), (90), (91), (92), (123), (124), (125) (126), (127),(128), (129), (130), (131), (132), (133), (134), (135), (136), (137),(138), (139) and (140).

The compounds of formulae (I), (Iab), (Ia), (Ib), (Ic), (Id), (Ie) and(If) can comprise one or more asymmetric carbon atoms. They can thusexist in the form of enantiomers or of diastereoisomers. Theseenantiomers, diastereoisomers and their mixtures, including the racemicmixtures, are encompassed within the scope of the present invention.

The compounds of the present invention can be prepared by conventionalmethods of organic synthesis practiced by those skilled in the art. Thegeneral reaction sequences outlined below represent a general methoduseful for preparing the compounds of the present invention and are notmeant to be limiting in scope or utility.

The compounds of general formula (I) can be prepared according to scheme1 below.

The synthesis is based on a coupling reaction starting from a halogenoaromatic compound of formula (III), wherein R₃, R₄, R₅ are as definedabove and X is a chlorine atom, an iodine atom or a bromine atom.

According to route (A1), the compound of formula (III) is placed in aprotic solvent such as tert-butanol. The compound of formula (II) isthen added in a molar ratio ranging from 1 to 1.5 with respect to thecompound of formula (III) in presence of an inorganic base, such asCs₂CO₃ or K₂CO₃ in a molar ratio ranging from 1 to 2, in the presence ofa diphosphine, such as Xantphos(4,5-Bis(diphenylphosphino)-9,9-dimethylxanthene) or X-Phos(2-Dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl) in an amountranging from 2 mol % to 10 mol % relative to the total amount ofcompound of formula (III), and in the presence of a catalyst, such asPd(OAc)₂ or Pd₂dba₃ in an amount ranging from 2 mol % and 10 mol %relative to the total amount of compound of formula (III). The reactionmixture can then be heated at a temperature ranging from 80 to 120° C.,for example at 90° C. and stirred for a time ranging from 15 to 25hours, for example during 20 hours, under inert gas and for exampleargon. The reaction mixture can be concentrated under reduced pressureand the residue can be diluted with an organic solvent such as ethylacetate. The organic phase can be washed with water, decanted and driedover magnesium sulphate. Finally, solid can be dried under vacuumovernight to give product (I).

According to route (A2), the compound of formula (III) is placed in apolar solvent such as dimethylformamide. The compound of formula (II) isthen added in a molar ratio ranging from 1 to 1.5 with respect to thecompound of formula (III) in presence of an inorganic base, such asCs₂CO₃ or K₂CO₃ in a molar ratio ranging from 1 to 2, in the presence ofa L-proline in an amount ranging from 10 mol % to 20 mol % relative tothe total amount of compound of formula (III), and in the presence of acatalyst, such as CuI in an amount ranging from 2 mol % and 10 mol %relative to the total amount of compound of formula (III). The reactionmixture can then be heated at a temperature ranging from 60 to 120° C.,for example at 80° C. and stirred for a time ranging from 24 to 72hours, for example during 48 hours, under inert gas and for exampleargon. The reaction mixture can be concentrated under reduced pressureand the residue can be diluted with an organic solvent such as ethylacetate. The organic phase can be washed with water, decanted and driedover magnesium sulphate. Finally, solid can be dried under vacuumovernight to give product (I).

The starting compounds of formula (II), (III) are available or can beprepared according to methods known to the person skilled in the art.

More particularly, compounds of formula (II) (i.e. respectively (IIa),(IIb) and (IIc)) when used to prepare compounds of formulae (Ia), (Ib)and (Ic) can be prepared according to scheme 2 below.

According to route (B), bromonitrobenzene is placed in a polar solventsuch as dimethylformamide. Vinylpyridine or styrene is then added in amolar ratio ranging from 1 to 1.5 with respect to the bromonitrobenzenein presence of an inorganic base such as sodium acetate or potassiumacetate in a molar ratio ranging from 1 to 3, in the presence of aphosphine such as triphenylphosphine in an amount ranging from 5% to 15mol % relative to the amount of bromonitrobenzene, and in the presenceof a catalyst such as Pd(OAc)₂ or Pd₂dba₃ in an amount ranging from 2%to 10% relative to the amount of bromonitrobenzene. The reaction mixturecan then be heated at a temperature ranging from 80 to 140° C., forexample at 135° C., and stirred for a time ranging from 15 to 30 hoursfor example 24 hours under inert gas for example argon. The reactionmixture can be concentrated under reduced pressure. The reaction mixturecan be concentrated under reduced pressure and the residue can bediluted with an organic solvent such as ethyl acetate. The organic phasecan be washed with water, decanted and dried over magnesium sulphate.Finally, solid can be dried under vacuum overnight to give product(IVa-c).

According to route (C), the compound of formula (IVa-c) and tin (II)chloride dihydrate in a ratio ranging from 3 to 8 equivalents are placedin a protic solvent such as ethanol. The reaction mixture can then beheated at a temperature ranging from 40 to 80° C., for example at 60° C.and stirred for a time ranging from 15 to 25 hours, for example during20 hours. The mixture can be poured into 1N NaOH aqueous solution andextracted with an organic solvent such as ethyl acetate. The organicphase can be decanted and dried over magnesium sulphate. Finally, solidcan be dried under vacuum overnight to give product (IIa-c).

More particularly, compounds of formula (II) (i.e. (IIc) when R10 istrifluoromethoxy) when used to prepare compounds of formula (Ic) can beprepared according to scheme 3 below.

According to route (D), methyl-triphenylphosphonium bromide in a ratioranging from 1.5 to 3 equivalents with respect to the nitrobenzaldehydeand potassium tert-butoxide in a ratio ranging from 1.5 to 3 equivalentswith respect to the nitrobenzaldehyde are placed in an aprotic solventsuch as toluene. The reaction mixture can then be heated at atemperature ranging from 50 to 110° C., for example at 70° C. andstirred for a time ranging from 15 to 50 minutes for example 30 minutesunder an inert atmosphere of argon. Nitrobenzaldehyde is then added. Thereaction mixture can then be heated at a temperature ranging from 70 to110° C., for example at 110° C. and for a time ranging from 1 to 4 hoursfor example 2 hours under an inert atmosphere of argon. Upon cooling toroom temperature, the reaction mixture can be diluted with water and theresulting solution can be extracted with an organic solvent such asethyl acetate. The organic phase can be decanted and dried overmagnesium sulphate. Finally, solid can be dried under vacuum overnightto give product (Vc).

The compounds of formula (II) (i.e. (IIf)), when used to preparecompounds of formula (If) can be prepared according to scheme 4 below.

According to route (E), bromonitrobenzene is placed in a protic solventsuch as tert-butanol. Benzamide is then added in a molar ratio rangingfrom 1 to 1.5 with respect to bromonitrobenzene in presence of aninorganic base, such as Cs₂CO₃ or K₂CO₃ in a molar ratio ranging from 1to 2, in the presence of a diphosphine, such as Xantphos(4,5-Bis(diphenylphosphino)-9,9-dimethylxanthene) or X-Phos(2-Dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl) in an amountranging from 2 mol % to 10 mol % relative to the total amount ofbromonitrobenzene, and in the presence of a catalyst, such as Pd(OAc)₂or Pd₂dba₃ in an amount ranging from 2 mol % and 10 mol % relative tothe total amount of bromonitrobenzene. The reaction mixture can then beheated at a temperature ranging from 80 to 120° C., for example at 90°C. and stirred for a time ranging from 15 to 25 hours, for exampleduring 20 hours, under inert gas and for example argon. The reactionmixture can be concentrated under reduced pressure and the residue canbe diluted with an organic solvent such as ethyl acetate. The organicphase can be washed with water, decanted and dried over magnesiumsulphate. Finally, solid can be dried under vacuum overnight to giveproduct (IVf).

According to route (F), the compound of formula (IVf) and 10% Pd/C in aratio ranging from 2% to 10% relative to the amount of benzamide areplaced in a protic solvent such as ethanol. The reaction mixture canthen be stirred for a time ranging from 2 to 18 hours for example 6hours under an atmosphere of H₂. The reaction mixture can then befiltered and the filtrate can be concentrated under reduced pressure togive product (IIf).

LEGENDS OF FIGURES

FIG. 1. Effect of drugs (40), (1) and (45) on p53 pathway in MCF7 celllines. (A) Analysis of p53 (total iso forms, blue panel) and its targetgene, Δ133p53 (Δ133 forms, purple panel), at mRNA level by RT-qPCR (B)Analysis of p53 expression at protein level using 4 differentantibodies. (C) Quantification of western blot (B) by densitometry usingImageJ software. (D) Analysis of p53-target genes expression by westernblot. (E) Quantification of western blot (D) by densitometry usingImageJ software.

FIG. 2. Effect of drugs (40), (1) and (45) on p53 pathway inMDA-MB-231-Luc-D3H2N cell lines maintained in normal medium. (A)Analysis of p53 (TA forms, blue panel) and its target gene, Δ133p53(Δ133 forms, purple panel), at mRNA level by RT-qPCR (B) Analysis of p53expression at protein level using 4 different antibodies. (C)Quantification of western blot (B) by densitometry using ImageJsoftware. (D) Analysis of p53-target genes expression by western blot.(E) Quantification of western blot (D) by densitometry using ImageJsoftware.

FIG. 3. Effect of drugs (40), (1) and (45) on p53 pathway inMDA-MB-231-Luc-D3H2N cell lines maintained in FCS-free medium. (A)Analysis of p53 (TA forms, blue panel) and its target gene, Δ133p53(Δ133 forms, purple panel), at mRNA level by RT-qPCR (B) Analysis of p53expression at protein level using 4 different antibodies. (C)Quantification of western blot (B) by densitometry using ImageJsoftware. (D) Analysis of p53-target genes expression by western blot.(E) Quantification of western blot (D) by densitometry using ImageJsoftware.

FIG. 4. Effect of drugs (40), (1) and (45) on p53 isoforms expression inMCF7 (A), MDA-MB-231-Luc-D3H2N cell lines maintained in normal medium(B) or in FCS-free medium (D). Analysis of C-terminal p53 isofromsproduced by alternative splicing (a, b and g forms) at mRNA level byRT-qPCR (left panel). An example of restoration of p53g-forms expressionis shown by nested RT-PCR (right panel).

The following example illustrates in detail the preparation of compounds(2), (1), (18), (9), (21), (127), (26), (40), (39), (45), (48), (49),(50), (52), (65) and (116)) according to the invention and thepharmacological data.

EXAMPLES Example 1: Compound (2) in Table I

According to route (B), 4-vinylpyridine (5.9 mL, 55 mmoles, 1.1 eq.) wasplaced in dimethylformamide (50 mL) with 1-bromo-3-nitrobenzene (10.1 g,50 mmoles, 1 eq.), NaOAc (8.2 g, 100 mmoles, 2 eq.), Pd(OAc)₂ (561 mg,2.5 mmoles, 5 mol %), PPh₃ (1.5 g, 6.0 mmoles, 12 mol %). The reactionmixture was heated at 135° C. and stirred for 24 hours under an inertatmosphere of argon. Upon cooling to room temperature, the reactionmixture was concentrated under reduced pressure and the resultingresidue was partitioned between ethyl acetate and water. Upondecantation, the aqueous phase was further extracted withdichloromethane. The organic phases were gathered, dried over MgSO₄,filtered and concentrated under reduced pressure. The resulting residuewas purified by column chromatography on silica gel to give(E)-4-[2-(3-nitrophenyl)vinyl]pyridine (6.0 g, 53%).

¹H NMR (300 MHz, CDCl₃) δ 8.63 (d, J=6.1 Hz, 2H), 8.41 (s, 1H), 8.17(dd, J=8.2, 2.1 Hz, 1H), 7.84 (d, J=7.8 Hz, 1H), 7.57 (t, J=8.0 Hz, 1H),7.40 (d, J=6.1 Hz, 2H), 7.34 (d, J=16.4 Hz, 1H), 7.16 (d, J=16.4 Hz,1H).

According to route (C), (E)-4-[2-(3-nitrophenyl)vinyl]pyridine (3.0 g,13.3 mmoles, 1 eq.) and tin (II) chloride dihydrate (15.0 g, 66.5mmoles, 5 eq.) were placed in EtOH (130 mL). The reaction mixture washeated at 60° C. and stirred for 16 hours under an inert atmosphere ofargon. The reaction mixture was then concentrated under reduced pressureand the resulting residue was diluted with ethyl acetate. The organicphase was washed with a 1N NaOH aqueous solution then with water, driedover MgSO₄, filtered and concentrated under reduced pressure to afford(E)-3-(2-pyridin-4-ylvinyl)phenylamine (1.9 g, 73%).

¹H NMR (300 MHz, CDCl₃) δ 8.57 (d, J=6.3 Hz, 2H), 7.35 (d, J=6.3 Hz,2H), 7.26-7.15 (m, 2H), 7.01-6.93 (m, 2H), 6.87 (s, 1H), 6.67 (dd,J=7.9, 2.3 Hz, 1H), 3.74 (br s, 2H).

N,N-diethylpropylenediamine (7 mL, 44 mmoles, 1.1 eq.) was placed in a3N NaOH aqueous solution (56 mL) and dichloromethane (24 mL) was addedto the solution. The reaction mixture was cooled down to 0° C. with anice bath and a solution of 3-bromobenzoyl chloride (5.3 mL, 40 mmoles, 1eq.) in dichloromethane (40 mL) was added dropwise. The reaction mixturewas then stirred at room temperature for 18 hours under an inertatmosphere of argon. Upon decantation, the organic phase was washed withwater, dried over MgSO₄, filtered and concentrated under reducedpressure to afford 3-bromo-N-(3-diethylamino-propyl)benzamide (12.5 g,99%).

¹H NMR (300 MHz, CDCl₃) δ 9.11 (br s, 1H), 7.90 (t, J=1.9 Hz, 1H), 7.72(dt, J=7.9, 1.2 Hz, 1H), 7.56 (ddd, J=7.9, 1.9, 1.0 Hz, 1H), 7.26 (t,J=7.9 Hz, 1H), 3.53 (dt, J=5.7, 4.6 Hz, 2H), 2.64-2.51 (m, 6H), 1.73(quint, J=5.7 Hz, 2H), 1.02 (t, J=7.2 Hz, 6H).

According to route (A1), a reaction mixture of3-bromo-N-(3-diethylamino-propyl)benzamide (1.4 g, 4.6 mmoles, 1 eq.),(E)-3-(2-pyridin-4-ylvinyl)phenylamine (1 g, 5.1 mmoles, 1.1 eq.),Pd₂(dba)₃ (211 mg, 0.23 mmole, 5 mol %), XPhos (219 mg, 0.46 mmole, 10mol %) and K₂CO₃ (2.6 g, 18.4 mmoles, 4 eq.) in t-BuOH (6.3 mL) washeated at 80° C. and stirred for 20 hours under an inert atmosphere ofargon. The reaction mixture was then concentrated under reduced pressureand the resulting residue was diluted with ethyl acetate. The organicphase was washed with water, dried over MgSO₄, filtered and concentratedunder reduced pressure. The resulting residue was purified by columnchromatography on alumina to give(E)-N-(3-diethylamino-propyl)-3-[3-(2-pyridin-4-ylvinyl)phenylamino]benzamide(2) (1.15 g, 58%).

¹H NMR (300 MHz, CDCl₃) δ 8.70 (s, 1H), 8.51 (d, J=4.9 Hz, 2H), 7.61 (s,1H), 7.30-7.14 (m, 8H), 7.08-7.00 (m, 2H), 6.89 (d, J=16.4 Hz, 1H), 6.58(s, 1H), 3.52-3.48 (m, 2H), 2.59-2.40 (m, 6H), 1.69-1.65 (m, 2H), 0.95(t, J=7.1 Hz, 6H).

¹³C NMR (75 MHz, CDCl₃) δ 167.1, 150.3, 144.7, 143.6, 143.5, 137.6,136.5, 133.3, 130.0, 129.4, 126.3, 121.1, 120.3, 120.2, 119.1, 118.8,117.1, 116.9, 53.4, 46.9, 41.4, 25.0, 11.6.

[M+H]⁺=429.2

Example 2: Compound (1) in Table I

According to route (B), 4-vinylpyridine (2.35 mL, 22 mmoles, 1.1 eq.)was placed in dimethylformamide (20 mL) with 1-bromo-3-nitrobenzene (4g, 20 mmoles, 1 eq.), NaOAc (3.3 g, 40 mmoles, 2 eq.), Pd(OAc)₂ (225 mg,1 mmole, 5 mol %), PPh₃ (629 mg, 2.4 mmoles, 12 mol %). The reactionmixture was heated at 135° C. and stirred for 24 hours under an inertatmosphere of argon. Upon cooling to room temperature, the reactionmixture was concentrated under reduced pressure and the resultingresidue was partitioned between ethyl acetate and water. Upondecantation, the organic phase was further washed with water, dried overMgSO₄, filtered and concentrated under reduced pressure to give(E)-4-[2-(3-nitrophenyl)vinyl]pyridine (2.9 g, 65%).

¹H NMR (300 MHz, CDCl₃) δ 8.64 (d, J=4.8 Hz, 2H), 8.41 (s, 1H), 8.17(dd, J=8.2, 1.1 Hz, 1H), 7.84 (d, J=7.8 Hz, 1H), 7.58 (t, J=8.0 Hz, 1H),7.40 (d, J=5.7 Hz, 2H), 7.34 (d, J=16.4 Hz, 1H), 7.16 (d, J=16.4 Hz,1H).

According to route (C), (E)-4-[2-(3-nitrophenyl)vinyl]pyridine (910 mg,4 mmoles, 1 eq.) and tin (II) chloride dihydrate (4.5 g, 20 mmoles, 5eq.) were placed in EtOH (40 mL). The reaction mixture was heated at 60°C. and stirred for 16 hours under an inert atmosphere of argon. Thereaction mixture was then concentrated under reduced pressure and theresulting residue was diluted with ethyl acetate. The organic phase waswashed with a 1N NaOH aqueous solution then with water, dried overMgSO₄, filtered and concentrated under reduced pressure to afford(E)-3-(2-pyridin-4-ylvinyl)phenylamine (763 mg, 97%).

¹H NMR (300 MHz, CDCl₃) δ 8.57 (d, J=5.9 Hz, 2H), 7.35 (d, J=5.9 Hz,2H), 7.26-7.14 (m, 2H), 7.01-6.92 (m, 2H), 6.87 (s, 1H), 6.67 (dd,J=7.9, 1.4 Hz, 1H), 3.74 (br s, 2H).

3-methyl-1-butanamine (2.5 mL, 22 mmoles, 1.1 eq.) was placed in a 3NNaOH aqueous solution (15 mL) and dichloromethane (5 mL) was added tothe solution. The reaction mixture was cooled down to 0° C. with an icebath and a solution of 3-bromobenzoyl chloride (2.6 mL, 20 mmoles, 1eq.) in dichloromethane (5 mL) was added dropwise. The reaction mixturewas then stirred at room temperature for 18 hours under an inertatmosphere of argon. Upon decantation, the organic phase was washed withwater, dried over MgSO₄, filtered and concentrated under reducedpressure to afford 3-bromo-N-(3-methylbutyl)benzamide (5.4 g, 100%).

¹H NMR (300 MHz, CDCl₃) δ 7.89 (s, 1H), 7.67 (d, J=7.9 Hz, 1H), 7.60 (d,J=7.9 Hz, 1H), 7.29 (d, J=7.9 Hz, 1H), 6.24 (br s, 1H), 3.45 (q, J=7.0Hz, 2H), 1.66 (heptuplet, J=6.6 Hz, 1H), 1.50 (q, J=7.0 Hz, 2H), 0.94(d, J=6.6 Hz, 6H).

According to route (A1), a reaction mixture of3-bromo-N-(3-methylbutyl)benzamide (373 mg, 1.38 mmole, 1 eq.),(E)-3-(2-pyridin-4-ylvinyl)phenylamine (300 mg, 1.53 mmole, 1.1 eq.),Pd₂(dba)₃ (63 mg, 69 μmoles, 5 mol %), XPhos (66 mg, 138 μmoles, 10 mol%) and K₂CO₃ (763 mg, 5.52 mmoles, 4 eq.) in t-BuOH (1.7 mL) was heatedat 90° C. and stirred for 20 hours under an inert atmosphere of argon.The reaction mixture was then concentrated under reduced pressure andthe resulting residue was diluted with ethyl acetate. The organic phasewas washed with water, dried over MgSO₄, filtered and concentrated underreduced pressure. The resulting residue was purified by columnchromatography on silica gel to give(E)-N-(3-methylbutyl)-3-[3-(2-pyridin-4-ylvinyl)phenylamino]benzamide(1) (333 mg, 63%).

¹H NMR (300 MHz, DMSO-d6) δ 8.52 (d, J=5.7 Hz, 2H), 8.39 (s, 1H),8.35-8.30 (m, 1H), 7.55 (d, J=6.0 Hz, 2H), 7.50 (d, J=16.7 Hz, 1H),7.32-7.27 (m, 5H), 7.20 (d, J=7.6 Hz, 2H), 7.15 (d, J=11.4 Hz, 1H), 7.05(d, J=8.2 Hz, 1H), 3.22 (q, J=6.9 Hz, 2H), 1.58 (heptuplet, J=6.6 Hz,1H), 1.37 (q, J=6.9 Hz, 2H), 0.87 (d, J=6.6 Hz, 6H).

¹³C NMR (75 MHz, DMSO-d6) δ 166.3, 150.0, 143.6, 143.4, 137.2, 136.1,133.3, 129.7, 129.1, 125.8, 120.9, 119.0, 118.8, 118.2, 117.5, 115.8,115.6, 38.1, 37.4, 25.3, 22.5.

[M+H]⁺=386.2

Example 3: Compound (18) in Table I

According to route (B), 4-vinylpyridine (6.4 mL, 59.9 mmoles, 1.1 eq.)was placed in dimethylformamide (55 mL) with 1-bromo-3-nitrobenzene (11g, 54.4 mmoles, 1 eq.), NaOAc (9 g, 108.8 mmoles, 2 eq.), Pd(OAc)₂ (610mg, 2.7 mmoles, 5 mol %), PPh₃ (1.4 g, 5.4 mmoles, 10 mol %). Thereaction mixture was heated at 135° C. and stirred for 24 hours under aninert atmosphere of argon. Upon cooling to room temperature, thereaction mixture was concentrated under reduced pressure and theresulting residue was partitioned between ethyl acetate and water. Upondecantation, the organic phase was further washed with water, dried overMgSO₄, filtered and concentrated under reduced pressure. The resultingresidue was purified by column chromatography on silica gel to give(E)-4-[2-(3-nitrophenyl)vinyl]pyridine (7.9 g, 64%).

¹H NMR (300 MHz, CDCl₃) δ 8.61 (d, J=5.0 Hz, 2H), 8.38 (s, 1H), 8.15 (d,J=8.1 Hz, 1H), 7.82 (d, J=7.6 Hz, 1H), 7.56 (t, J=7.9 Hz, 1H), 7.38 (d,J=5.0 Hz, 2H), 7.32 (d, J=16.4 Hz, 1H), 7.14 (d, J=16.4 Hz, 1H).

According to route (C), (E)-4-[2-(3-nitrophenyl)vinyl]pyridine (7.9 g,35 mmoles, 1 eq.) and tin (II) chloride dihydrate (39 g, 175 mmoles, 5eq.) were placed in EtOH (350 mL). The reaction mixture was heated at60° C. and stirred for 48 hours under an inert atmosphere of argon. Thereaction mixture was then concentrated under reduced pressure and theresulting residue was diluted with ethyl acetate. The organic phase waswashed with a 1N NaOH aqueous solution then with water, dried overMgSO₄, filtered and concentrated under reduced pressure to afford(E)-3-(2-pyridin-4-ylvinyl)phenylamine (5.2 g, 76%).

¹H NMR (300 MHz, CDCl₃) δ 8.57 (d, J=6.3 Hz, 2H), 7.35 (d, J=6.3 Hz,2H), 7.30-7.16 (m, 2H), 7.03-6.91 (m, 2H), 6.86 (s, 1H), 6.72-6.60 (m,1H), 3.74 (br s, 2H).

3-bromobenzenesulfonyl chloride (0.56 mL, 3.9 mmoles, 1 eq.) andN,N-diisopropylethylamine (1.02 mL, 5.9 mmoles, 1.5 eq.) were placed inanhydrous dichloromethane (20 mL). The reaction mixture was cooled downto 0° C. with an ice bath and N,N-diethylpropylenediamine (1.23 mL, 7.8mmoles, 2 eq.) was added dropwise. The reaction mixture was then stirredat 0° C. for 2 hours under an inert atmosphere of argon. The mixture waswashed with saturated aqueous solutions of NH₄Cl and then NaCl. Theaqueous phases were extracted with dichloromethane. The organic phaseswere gathered, dried over MgSO₄, filtered and concentrated under reducedpressure to afford 3-bromo-N-(3-diethylaminopropyl)benzenesulfonamide(524 mg, 38%).

¹H NMR (300 MHz, CDCl₃) δ 7.98 (s, 1H), 7.78 (d, J=7.9 Hz, 1H), 7.66 (d,J=7.9 Hz, 1H), 7.37 (t, J=7.9 Hz, 1H), 3.12-2.96 (m, 2H), 2.67-2.43 (m,6H), 1.77-1.58 (m, 2H), 1.06 (t, J=7.1 Hz, 6H).

According to route (A1), a reaction mixture of3-bromo-N-(3-diethylaminopropyl)benzenesulfonamide (175 mg, 0.50 mmole,1 eq.), (E)-3-(2-pyridin-4-ylvinyl)phenylamine (108 mg, 0.55 mmole, 1.1eq.), Pd₂(dba)₃ (23 mg, 0.025 mmole, 5 mol %), XPhos (24 mg, 0.05 mmole,10 mol %) and K₂CO₃ (276 mg, 2 mmoles, 4 eq.) in t-BuOH (2 mL) washeated at 90° C. and stirred for 20 hours under an inert atmosphere ofargon. The reaction mixture was then concentrated under reduced pressureand the resulting residue was diluted with ethyl acetate. The organicphase was washed with water, dried over MgSO₄, filtered and concentratedunder reduced pressure. The resulting residue was purified by columnchromatography on silica gel to giveN-(3-diethylaminopropyl)-3-[3-(2-pyridin-4-ylvinyl)phenylamino]benzenesulfonamide(18) (100 mg, 43%).

¹H NMR (300 MHz, CDCl₃) δ 8.55 (d, J=5.8 Hz, 2H), 7.58 (s, 1H),7.39-7.32 (m, 4H), 7.31-7.19 (m, 4H), 7.15 (d, J=7.7 Hz, 1H), 7.10 (d,J=7.9 Hz, 1H), 6.98 (d, J=16.3 Hz, 1H), 6.64 (d, J=8.4 Hz, 1H),3.15-3.00 (m, 2H), 2.52-2.35 (m, 6H), 1.68-1.53 (m, 2H), 0.96 (t, J=7.1Hz, 6H).

[M+H]⁺=465.2

Example 4: Compound (9) in Table I

According to route (B), 4-vinylpyridine (5.9 mL, 55 mmoles, 1.1 eq.) wasplaced in dimethylformamide (50 mL) with 1-bromo-3-nitrobenzene (10.1 g,50 mmoles, 1 eq.), NaOAc (8.2 g, 100 mmoles, 2 eq.), Pd(OAc)₂ (561 mg,2.5 mmoles, 5 mol %), PPh₃ (1.5 g, 6.0 mmoles, 12 mol %). The reactionmixture was heated at 135° C. and stirred for 24 hours under an inertatmosphere of argon. Upon cooling to room temperature, the reactionmixture was concentrated under reduced pressure and the resultingresidue was partitioned between ethyl acetate and water. Upondecantation, the aqueous phase was further extracted withdichloromethane. The organic phases were gathered, dried over MgSO₄,filtered and concentrated under reduced pressure. The resulting residuewas purified by column chromatography on silica gel to give(E)-4-[2-(3-nitrophenyl)vinyl]pyridine (6.0 g, 53%).

¹H NMR (300 MHz, CDCl₃) δ 8.63 (d, J=6.1 Hz, 2H), 8.41 (s, 1H), 8.17(dd, J=8.2, 2.1 Hz, 1H), 7.84 (d, J=7.8 Hz, 1H), 7.57 (t, J=8.0 Hz, 1H),7.40 (d, J=6.1 Hz, 2H), 7.34 (d, J=16.4 Hz, 1H), 7.16 (d, J=16.4 Hz,1H).

According to route (C), (E)-4-[2-(3-nitrophenyl)vinyl]pyridine (3.0 g,13.3 mmoles, 1 eq.) and tin (II) chloride dihydrate (15.0 g, 66.5mmoles, 5 eq.) were placed in EtOH (130 mL). The reaction mixture washeated at 60° C. and stirred for 16 hours under an inert atmosphere ofargon. The reaction mixture was then concentrated under reduced pressureand the resulting residue was diluted with ethyl acetate. The organicphase was washed with a 1N NaOH aqueous solution then with water, driedover MgSO₄, filtered and concentrated under reduced pressure to afford(E)-3-(2-pyridin-4-ylvinyl)phenylamine (1.9 g, 73%).

¹H NMR (300 MHz, CDCl₃) δ 8.57 (d, J=6.3 Hz, 2H), 7.35 (d, J=6.3 Hz,2H), 7.26-7.15 (m, 2H), 7.01-6.93 (m, 2H), 6.87 (s, 1H), 6.67 (dd,J=7.9, 2.3 Hz, 1H), 3.74 (br s, 2H).

NH4OH solution (5.8 mL) was placed in water (24 mL) at −10° C.3-bromobenzoyl chloride (10 mmol, 1.3 mL) was placed in tetrahydrofurane(8 mL) and added to the aqueous solution. The mixture was stirred for 1hour at −10° C. The reaction mixture was filtered and further washedwith water to give 3-beomo-benzamide (1.7 g, 85%).

¹H NMR (300 MHz, MeOD) δ 8.04 (s, 1H), 7.84 (d, J=7.0 Hz, 1H), 7.70 (d,J=6.6 Hz, 1H), 7.40-7.38 (t, J=6.8 Hz, 1H).

According to route (A1), a reaction mixture of 3-bromo-benzamide (99 mg,0.5 mmoles, 1 eq.), (E)-3-(2-pyridin-4-ylvinyl)phenylamine (108 mg, 0.55mmoles, 1.1 eq.), Pd₂(dba)₃ (23 mg, 0.025 mmole, 5 mol %), XPhos (143mg, 0.05 mmole, 10 mol %) and K₂CO₃ (276 mg, 2.0 mmoles, 4 eq.) int-BuOH (2.0 mL) was heated at 90° C. and stirred for 20 hours under aninert atmosphere of argon. The reaction mixture was then concentratedunder reduced pressure and the resulting residue was partitioned betweenethyl acetate and water. Upon decantation, the aqueous phase was furtherextracted with dichloromethane. The organic phases were gathered, driedover MgSO₄, filtered and concentrated under reduced pressure. Theresulting residue was purified by column chromatography on silica gel togive 3-[3-(2-pyridin-4-ylvinyl)phenylamino]benzamide (9) (100 mg, 63%).

¹H NMR (300 MHz, MeOD) δ 8.46 (d, J=6.2 Hz, 1H), 7.62 (s, 1H), 7.57 (d,J=6.1 Hz, 1H), 7.46 (d, J=16.1 Hz, 1H), 7.38-7.30 (m, 2H), 7.28 (d,J=8.1 Hz, 1H), 7.18 (d, J=7.8 Hz, 1H), 7.16-7.07 (m, 1H).

[M+H]⁺=314.0

Example 5: Compound (21) in Table I

According to route (B), 4-vinylpyridine (5.9 mL, 55 mmoles, 1.1 eq.) wasplaced in dimethylformamide (50 mL) with 1-bromo-3-nitrobenzene (10.1 g,50 mmoles, 1 eq.), NaOAc (8.2 g, 100 mmoles, 2 eq.), Pd(OAc)₂ (561 mg,2.5 mmoles, 5 mol %), PPh₃ (1.5 g, 6.0 mmoles, 12 mol %). The reactionmixture was heated at 135° C. and stirred for 24 hours under an inertatmosphere of argon. Upon cooling to room temperature, the reactionmixture was concentrated under reduced pressure and the resultingresidue was partitioned between ethyl acetate and water. Upondecantation, the aqueous phase was further extracted withdichloromethane. The organic phases were gathered, dried over MgSO₄,filtered and concentrated under reduced pressure. The resulting residuewas purified by column chromatography on silica gel to give(E)-4-[2-(3-nitrophenyl)vinyl]pyridine (6.0 g, 53%).

¹H NMR (300 MHz, CDCl₃) δ 8.63 (d, J=6.1 Hz, 2H), 8.41 (s, 1H), 8.17(dd, J=8.2, 2.1 Hz, 1H), 7.84 (d, J=7.8 Hz, 1H), 7.57 (t, J=8.0 Hz, 1H),7.40 (d, J=6.1 Hz, 2H), 7.34 (d, J=16.4 Hz, 1H), 7.16 (d, J=16.4 Hz,1H).

According to route (C), (E)-4-[2-(3-nitrophenyl)vinyl]pyridine (3.0 g,13.3 mmoles, 1 eq.) and tin (II) chloride dihydrate (15.0 g, 66.5mmoles, 5 eq.) were placed in EtOH (130 mL). The reaction mixture washeated at 60° C. and stirred for 16 hours under an inert atmosphere ofargon. The reaction mixture was then concentrated under reduced pressureand the resulting residue was diluted with ethyl acetate. The organicphase was washed with a 1N NaOH aqueous solution then with water, driedover MgSO₄, filtered and concentrated under reduced pressure to afford(E)-3-(2-pyridin-4-ylvinyl)phenylamine (1.9 g, 73%).

¹H NMR (300 MHz, CDCl₃) δ 8.57 (d, J=6.3 Hz, 2H), 7.35 (d, J=6.3 Hz,2H), 7.26-7.15 (m, 2H), 7.01-6.93 (m, 2H), 6.87 (s, 1H), 6.67 (dd,J=7.9, 2.3 Hz, 1H), 3.74 (br s, 2H).

3-(4-methyl-piperazinyl)-propylamine (1.9 mL, 11 mmoles, 1.1 eq.) wasplaced in a 3N NaOH aqueous solution (14 mL) and dichloromethane (2 mL)was added to the solution. The reaction mixture was cooled down to 0° C.with an ice bath and a solution of 3-bromobenzoyl chloride (1.9 mL, 10mmoles, 1 eq.) in dichloromethane (14 mL) was added dropwise. Thereaction mixture was then stirred at room temperature for 18 hours underan inert atmosphere of argon. Upon decantation, the organic phase waswashed with water, dried over MgSO₄, filtered and concentrated underreduced pressure to afford3-bromo-N-[3-(4-methyl-piperazinyl)]-propylamine (2.8 g, 85%).

¹H NMR (300 MHz, CDCl₃) δ 8.59 (s, 1H), 7.90 (s, 1H), 7.79 (d, J=8.4 Hz,1H), 7.59 (d, J=7.9 Hz, 1H), 7.28 (t, J=8.2 Hz, 1H), 3.54 (d, J=4.7 Hz,2H), 2.63-2.50 (m, 4H), 2.46 (broad s, 4H), 2.30 (s, 5H), 1.89-1.54 (m,2H).

According to route (A1), a reaction mixture of3-bromo-N-[3-(4-methyl-piperazinyl)]-propylamine (169 mg, 0.50 mmoles, 1eq.), (E)-3-(2-pyridin-4-ylvinyl)phenylamine (108 mg, 0.55 mmoles, 1.1eq.), Pd₂(dba)₃ (23 mg, 0.025 mmole, 5 mol %), XPhos (24 mg, 0.05 mmole,10 mol %) and K₂CO₃ (276 mg, 2.0 mmoles, 4 eq.) in t-BuOH (2 mL) washeated at 80° C. and stirred for 20 hours under an inert atmosphere ofargon. The reaction mixture was then concentrated under reduced pressureand the resulting residue was diluted with ethyl acetate. The organicphase was washed with water, dried over MgSO₄, filtered and concentratedunder reduced pressure. The resulting residue was purified by columnchromatography on alumina to give(E)-[3-(4-methyl-piperazinyl)-propyl]-3-[3-(2-pyridin-4-ylvinyl)-phenylamino]-benzamide(21) (115 mg, 51%).

¹H NMR (300 MHz, CDCl₃) δ 8.49 (d, J=3.6 Hz, 2H), 8.23 (s, 1H), 7.56 (s,1H), 7.39-7.11 (m, 7H), 7.14-6.98 (m, 3H), 6.90 (d, J=15.8 Hz, 1H), 6.53(s, 1H), 3.49 (s, 2H), 3.23 (s, 2H), 2.2.5-2.30 (m, 8H), 2.20 (s, 3H),1.75-1.70 (m, 2H).

¹³C NMR (75 MHz, CDCl3) δ 167.47, 150.25, 144.65, 143.65, 143.38,137.49, 136.43, 133.24, 129.95, 129.35, 126.26, 121.02, 120.13, 119.99,119.17, 118.56, 117.07, 116.68, 58.36, 55.12, 53.37, 46.13, 40.93,24.40.

[M+1-1]⁺=456.2

Example 6: Compound (127) in Table I

According to route (B), 4-vinylpyridine (5.9 mL, 55 mmoles, 1.1 eq.) wasplaced in dimethylformamide (50 mL) with 1-bromo-3-nitrobenzene (10.1 g,50 mmoles, 1 eq.), NaOAc (8.2 g, 100 mmoles, 2 eq.), Pd(OAc)₂ (561 mg,2.5 mmoles, 5 mol %), PPh₃ (1.5 g, 6.0 mmoles, 12 mol %). The reactionmixture was heated at 135° C. and stirred for 24 hours under an inertatmosphere of argon. Upon cooling to room temperature, the reactionmixture was concentrated under reduced pressure and the resultingresidue was partitioned between ethyl acetate and water. Upondecantation, the aqueous phase was further extracted withdichloromethane. The organic phases were gathered, dried over MgSO₄,filtered and concentrated under reduced pressure. The resulting residuewas purified by column chromatography on silica gel to give(E)-4-[2-(3-nitrophenyl)vinyl]pyridine (6.0 g, 53%).

¹H NMR (300 MHz, CDCl₃) δ 8.63 (d, J=6.1 Hz, 2H), 8.41 (s, 1H), 8.17(dd, J=8.2, 2.1 Hz, 1H), 7.84 (d, J=7.8 Hz, 1H), 7.57 (t, J=8.0 Hz, 1H),7.40 (d, J=6.1 Hz, 2H), 7.34 (d, J=16.4 Hz, 1H), 7.16 (d, J=16.4 Hz,1H).

According to route (C), (E)-4-[2-(3-nitrophenyl)vinyl]pyridine (3.0 g,13.3 mmoles, 1 eq.) and tin (II) chloride dihydrate (15.0 g, 66.5mmoles, 5 eq.) were placed in EtOH (130 mL). The reaction mixture washeated at 60° C. and stirred for 16 hours under an inert atmosphere ofargon. The reaction mixture was then concentrated under reduced pressureand the resulting residue was diluted with ethyl acetate. The organicphase was washed with a 1N NaOH aqueous solution then with water, driedover MgSO₄, filtered and concentrated under reduced pressure to afford(E)-3-(2-pyridin-4-ylvinyl)phenylamine (1.9 g, 73%).

¹H NMR (300 MHz, CDCl₃) δ 8.57 (d, J=6.3 Hz, 2H), 7.35 (d, J=6.3 Hz,2H), 7.26-7.15 (m, 2H), 7.01-6.93 (m, 2H), 6.87 (s, 1H), 6.67 (dd,J=7.9, 2.3 Hz, 1H), 3.74 (br s, 2H).

According to route (A1), a reaction mixture of3-bromo-benzenesulfonamide (118 mg, 0.50 mmoles, 1 eq.),(E)-3-(2-pyridin-4-ylvinyl)phenylamine (108 mg, 0.55 mmoles, 1.1 eq.),Pd₂(dba)₃ (23 mg, 0.025 mmole, 5 mol %), XPhos (24 mg, 0.05 mmole, 10mol %) and K₂CO₃ (276 mg, 2.0 mmoles, 4 eq.) in t-BuOH (2 mL) was heatedat 80° C. and stirred for 20 hours under an inert atmosphere of argon.The reaction mixture was then concentrated under reduced pressure andthe resulting residue was diluted with ethyl acetate. The organic phasewas washed with water, dried over MgSO₄, filtered and concentrated underreduced pressure. The resulting residue was purified by columnchromatography on alumina to give(E)-3-[3-(2-pyridin-4-ylvinyl)phenylamino]-benzenesulfonamide (127) (89mg, 51%).

¹H NMR (300 MHz, MeOD) δ 8.43 (s, 2H), 7.67 (s, 1H), 7.53 (d, J=5.2 Hz,2H), 7.42 (d, J=16.5 Hz, 1H), 7.38-7.13 (m, 8H), 7.13-7.07 (m, 1H), 7.09(d, J=16.5 Hz, 1H).

[M+H]⁺=352.1

Example 7: Compound (26) in Table I

According to route (B), 2-vinylpyridine (1.18 mL, 11 mmoles, 1.1 eq.)was placed in dimethylformamide (10 mL) with 1-bromo-3-nitrobenzene(2.02 g, 10 mmoles, 1 eq.), NaOAc (1.64 g, 20 mmoles, 2 eq.), Pd(OAc)₂(112 mg, 0.5 mmole, 5 mol %), PPh₃ (315 mg, 1.2 mmole, 12 mol %). Thereaction mixture was heated at 135° C. and stirred for 24 hours under aninert atmosphere of argon. Upon cooling to room temperature, thereaction mixture was concentrated under reduced pressure and theresulting residue was partitioned between ethyl acetate and water. Upondecantation, the organic phase was further washed with water, dried overMgSO₄, filtered and concentrated under reduced pressure. The resultingresidue was purified by column chromatography on silica gel to give(E)-2-[2-(3-nitrophenyl)vinyl]pyridine (1.0 g, 44%).

¹H NMR (300 MHz, CDCl₃) δ 8.64 (d, J=4.6 Hz, 1H), 8.44 (s, 1H), 8.14(ddd, J=8.2, 2.1, 0.9 Hz, 1H), 7.86 (d, J=7.7 Hz, 1H), 7.77-7.65 (m,2H), 7.55 (t, J=8.0 Hz, 1H), 7.41 (d, J=7.9 Hz, 1H), 7.28 (d, J=16.0 Hz,1H), 7.22 (ddd, J=7.5, 4.8, 1.0 Hz, 1H).

According to route (C), (E)-2-[2-(3-nitrophenyl)vinyl]pyridine (1.0 g,4.4 mmoles, 1 eq.) and tin (II) chloride dihydrate (5.0 g, 22.1 mmoles,5 eq.) were placed in EtOH (44 mL). The reaction mixture was heated at60° C. and stirred for 16 hours under an inert atmosphere of argon. Thereaction mixture was then concentrated under reduced pressure and theresulting residue was diluted with ethyl acetate. The organic phase waswashed with a 1N NaOH aqueous solution then with water, dried overMgSO₄, filtered and concentrated under reduced pressure to afford(E)-3-(2-pyridin-2-ylvinyl)phenylamine (816 mg, 94%).

¹H NMR (300 MHz, CDCl₃) δ 8.58 (dd, J=4.8, 0.7 Hz, 1H), 7.60 (td, J=7.7,1.8 Hz, 1H), 7.53 (d, J=16.1 Hz, 1H), 7.33 (d, J=7.9 Hz, 1H), 7.18-7.05(m, 3H), 6.97 (d, J=7.7 Hz, 1H), 6.86 (s, 1H), 6.61 (dd, J=7.9, 1.4 Hz,1H), 3.76 (br s, 1H).

Pent-4-yn-1-ol (5 g, 59 mmoles, 1.0 eq.) was placed in adimethylsulfoxide (65 mL) and water (5 mL) solution, together with1-bromo-4-iodobenzene (16.81 g, 59 mmoles, 1 eq.), NaN₃ (4.64 g, 71.3mmoles, 1.2 eq.), L-Proline (1.53 g, 11.8 mmoles, 0.2 eq.), Na₂CO₃ (1.26g, 11.8 mmoles, 0.2 eq.), sodium ascorbate (3.57 g, 23.7 mmoles, 0.4eq.). CuSO₄.5H₂O (4.7 g, 11.8 mmoles, 0.2 eq.) was added and thereaction mixture was heated at 65° C. and stirred for 16 hours under aninert atmosphere of argon. Upon cooling to room temperature, thereaction mixture was further stirred for 24 hours and then partitionedbetween a NH₄OH aqueous solution and ethyl acetate. Upon decantation,the aqueous phase was further extracted with ethyl acetate. The organicphases were gathered, washed with a NaCl aqueous solution, dried overMgSO₄, filtered and concentrated under reduced pressure to afford3-[1-(4-bromophenyl)-1H-[1,2,3]triazol-4-yl]propan-1-ol (10.36 g, 53%).

¹H NMR (300 MHz, DMSO-d6) δ 8.61 (s, 1H), 7.86 (d, J=8.9 Hz, 1H), 7.79(d, J=8.9 Hz, 1H), 4.52 (t, J=5.2 Hz, 1H), 3.48 (q, J=5.2 Hz, 2H), 2.73(t, J=7.5 Hz, 2H), 1.88-1.74 (quint, J=7.5 Hz, 2H).

According to route (A1), a reaction mixture of3-[1-(4-bromophenyl)-1H-[1,2,3]triazol-4-yl]propan-1-ol (846 mg, 3.0mmoles, 1 eq.), (E)-3-(2-pyridin-2-ylvinyl)phenylamine (816 mg, 4.2mmoles, 1.4 eq.), Pd₂(dba)₃ (137 mg, 0.15 mmole, 5 mol %), XPhos (143mg, 0.30 mmole, 10 mol %) and K₂CO₃ (1.66 g, 12.0 mmoles, 4 eq.) int-BuOH (3.7 mL) was heated at 90° C. and stirred for 20 hours under aninert atmosphere of argon. The reaction mixture was then concentratedunder reduced pressure and the resulting residue was partitioned betweenethyl acetate and water. Upon decantation, the aqueous phase was furtherextracted with dichloromethane. The organic phases were gathered, driedover MgSO₄, filtered and concentrated under reduced pressure. Theresulting residue was purified by column chromatography on silica gel togive3-(1-{4-[3-(2-pyridin-2-ylvinyl)phenylamino]phenyl}-1H-[1,2,3]triazol-4-yl)propan-1-ol(26) (836 mg, 70%).

¹H NMR (300 MHz, CDCl₃) δ 8.51 (d, J=4.5, 1H), 7.61 (s, 1H), 7.56 (d,J=7.6, 1H), 7.48 (d, J=16.1, 1H), 7.41 (d, J=8.7, 2H), 7.30 (d, J=7.8,1H), 7.21 (s, 1H), 7.17 (d, J=7.7, 1H), 7.12-7.01 (m, 5H), 6.97 (d,J=7.8, 1H), 6.87 (s, 1H), 4.19 (s, 1H), 3.70 (t, J=6.2, 2H), 2.84 (t,J=7.4, 2H), 2.00 (quint, J=6.7, 2H).

¹³C NMR (75 MHz, CDCl₃) δ 155.6, 149.8, 148.2, 144.0, 142.7, 138.1,136.9, 132.7, 130.3, 130.0, 128.4, 122.4, 122.1, 121.0, 119.5, 118.9,117.7, 117.3, 61.9, 32.2, 22.3.

[M+H]⁺=398

Example 8: Compound (40) in Table I

According to route (B), 1-methoxy-4-vinylbenzene (2.9 mL, 22 mmoles, 1.1eq.) was placed in dimethylformamide (20 mL) with 1-bromo-3-nitrobenzene(4 g, 20 mmoles, 1 eq.), NaOAc (3.3 g, 40 mmoles, 2 eq.), Pd(OAc)₂ (225mg, 1 mmole, 5 mol %), PPh₃ (629 mg, 2.4 mmoles, 12 mol %). The reactionmixture was heated at 135° C. and stirred for 24 hours under an inertatmosphere of argon. Upon cooling to room temperature, the reactionmixture was concentrated under reduced pressure and the resultingresidue was partitioned between ethyl acetate and water. Upondecantation, the aqueous phase was further extracted withdichloromethane. The organic phases were gathered, dried over MgSO₄,filtered and concentrated under reduced pressure. The resulting residuewas purified by column chromatography on silica gel to afford(E)-1-nitro-3-(4-(methoxy)styryl)benzene (2.3 g, 45%).

¹H NMR (300 MHz, CDCl₃) δ 8.35 (s, 1H), 8.07 (d, J=8.8 Hz, 1H), 7.77 (d,J=7.7 Hz, 1H), 7.56-7.43 (m, 3H), 7.20 (d, J=16.4 Hz, 1H), 7.01 (d,J=16.4 Hz, 1H), 6.94 (d, J=7.9 Hz, 2H), 3.86 (s, 3H).

According to route (C), (E)-1-nitro-3-(4-(methoxy)styryl)benzene (1.5 g,5.8 mmoles, 1 eq.) and tin (II) chloride dihydrate (6.6 g, 29.4 mmoles,5 eq.) were placed in EtOH (58 mL). The reaction mixture was heated at60° C. and stirred for 48 hours under an inert atmosphere of argon. Thereaction mixture was then concentrated under reduced pressure and theresulting residue was diluted with ethyl acetate. The organic phase waswashed with a 1N NaOH aqueous solution, dried over MgSO₄, filtered andconcentrated under reduced pressure to afford(E)-1-nitro-3-(4-(methoxy)styryl)aniline (1.3 g, 99%).

¹H NMR (300 MHz, CDCl₃) δ 7.46 (d, J=8.7 Hz, 2H), 7.17 (t, J=7.8 Hz,1H), 7.05 (d, J=16.3 Hz, 1H), 6.97-6.88 (m, 4H), 6.84 (s, 1H), 6.60 (dd,J=7.9, 2.1 Hz, 1H), 3.84 (s, 3H), 3.68 (br s, 2H).

N,N-diethylpropylenediamine (7 mL, 44 mmoles, 1.1 eq.) was placed in a3N NaOH aqueous solution (56 mL) and dichloromethane (24 mL) was addedto the solution. The reaction mixture was cooled down to 0° C. with anice bath and a solution of 3-bromobenzoyl chloride (5.3 mL, 40 mmoles, 1eq.) in dichloromethane (40 mL) was added dropwise. The reaction mixturewas then stirred at room temperature for 18 hours under an inertatmosphere of argon. Upon decantation, the organic phase was washed withwater, dried over MgSO₄, filtered and concentrated under reducedpressure to afford 3-bromo-N-(3-diethylamino-propyl)benzamide (12.5 g,99%).

¹H NMR (300 MHz, CDCl₃) δ 9.11 (br s, 1H), 7.90 (t, J=1.9 Hz, 1H), 7.72(dt, J=7.9, 1.2 Hz, 1H), 7.56 (ddd, J=7.9, 1.9, 1.0 Hz, 1H), 7.26 (t,J=7.9 Hz, 1H), 3.53 (dt, J=5.7, 4.6 Hz, 2H), 2.64-2.51 (m, 6H), 1.73(quint, J=5.7 Hz, 2H), 1.02 (t, J=7.2 Hz, 6H).

According to route (A1), a reaction mixture of3-bromo-N-(3-diethylamino-propyl)benzamide (1.2 g, 4 mmoles, 1 eq.),(E)-1-nitro-3-(4-(methoxy)styryl)aniline (1 g, 4.4 mmoles, 1.1 eq.),Pd₂(dba)₃ (183 mg, 0.2 mmole, 5 mol %), XPhos (190 mg, 0.4 mmole, 10 mol%) and K₂CO₃ (2.2 g, 16 mmoles, 4 eq.) in t-BuOH (5 mL) was heated at90° C. and stirred for 20 hours under an inert atmosphere of argon. Thereaction mixture was then concentrated under reduced pressure and theresulting residue was diluted with ethyl acetate. The organic phase waswashed with water, dried over MgSO₄, filtered and concentrated underreduced pressure. The resulting residue was purified by columnchromatography on silica gel to give(E)-N-(3-diethylamino-propyl)-3-((3-(4-(methoxy)styryl)phenylamino)benzamide(40) (1 g, 55%).

¹H NMR (300 MHz, CDCl₃) δ 8.59 (s, 1H), 7.52 (s, 1H), 7.43 (d, J=8.7 Hz,2H), 7.27 (d, J=7.0 Hz, 2H), 7.23 (d, J=7.3 Hz, 2H), 7.18 (s, 1H),7.13-6.92 (m, 4H), 6.89 (d, J=8.7 Hz, 2H), 3.82 (s, 3H), 3.59-3.47 (m,2H), 2.62-2.46 (m, 6H), 1.72 (quint, J=5.7 Hz, 2H), 0.99 (t, J=7.2 Hz,6H).

¹³C NMR (75 MHz, CDCl₃) δ 167.2, 159.5, 144.0, 143.2, 139.1, 136.4,130.2, 129.8, 129.4, 128.6, 127.9, 119.8, 118.9, 117.6, 116.6, 114.3,55.5, 53.3, 46.9, 41.3, 25.1, 11.6.

[M+H]⁺=458.3

Example 9: Compound (39) in Table I

According to route (D), methyl-triphenylphosphonium bromide (3.6 g, 10mmoles, 2 eq.) was placed in dry toluene (17 mL) with potassiumtert-butoxide (1.1 g, 10 mmoles, 2 eq.). The reaction mixture was heatedat 70° C. and stirred for 30 minutes under an inert atmosphere of argon.3-Nitrobenzaldehyde (756 mg, 5 mmoles, 1 eq.) was then added. Thereaction mixture was heated at 110° C. and stirred for 2 hours under aninert atmosphere of argon. Upon cooling to room temperature, thereaction mixture was diluted with water and the resulting solution wasextracted with ethyl acetate. The organic phase was dried over MgSO₄,filtered and concentrated under reduced pressure. The resulting residuewas purified by column chromatography on silica gel to afford1-nitro-3-vinylbenzene (661 mg, 88%).

¹H NMR (300 MHz, CDCl₃) δ 8.20 (s, 1H), 8.06 (s, 1H), 7.68 (s, 1H), 7.46(s, 1H), 6.83-6.66 (m, 1H), 5.87 (d, J=17.5 Hz, 1H), 5.41 (d, J=10.7 Hz,1H).

According to route (B), 1-nitro-3-vinylbenzene (661 mg, 4.4 mmoles, 1.1eq.) was placed in dimethylformamide (4 mL) with 1-bromo-3(trifluoromethoxy)benzene (600 μL, 4 mmoles, 1 eq.), NaOAc (661 mg, 8mmoles, 2 eq.), Pd(OAc)₂ (45 mg, 0.2 mmole, 5 mol %), PPh₃ (105 mg, 0.4mmole, 10 mol %). The reaction mixture was heated at 135° C. and stirredfor 10 hours under an inert atmosphere of argon. Upon cooling to roomtemperature, the reaction mixture was concentrated under reducedpressure and the resulting residue was partitioned between ethyl acetateand water. Upon decantation, the aqueous phase was further extractedwith dichloromethane. The organic phases were gathered, dried overMgSO₄, filtered and concentrated under reduced pressure. The resultingresidue was purified by column chromatography on silica gel to afford(E)-1-nitro-3-(4-(methoxy)styryl)benzene (355 mg, 28%).

¹H NMR (300 MHz, CDCl₃) δ 8.37 (s, 1H), 8.13 (d, J=8.4 Hz, 1H), 7.81 (d,J=7.7 Hz, 1H), 7.54 (t, J=7.9 Hz, 1H), 7.49-7.34 (m, 3H), 7.20-7.00 (m,3H).

According to route (C),(E)-1-nitro-3-(3-(trifluoromethoxy)styryl)benzene (355 mg, 1.15 mmole, 1eq.) and tin (II) chloride dihydrate (1.3 g, 5.75 mmoles, 5 eq.) wereplaced in EtOH (11.5 mL). The reaction mixture was heated at 60° C. andstirred for 10 hours under an inert atmosphere of argon. The reactionmixture was then concentrated under reduced pressure and the resultingresidue was diluted with ethyl acetate. The organic phase was washedwith a 1N NaOH aqueous solution, dried over MgSO₄, filtered andconcentrated under reduced pressure to afford(E)-3-(3-(trifluoromethoxy)styryl)aniline (304 mg, 95%).

¹H NMR (300 MHz, CDCl₃) δ 7.45-7.34 (m, 3H), 7.20 (t, J=7.8 Hz, 1H),7.13 (d, J=8.5 Hz, 1H), 7.05 (s, 2H), 6.96 (d, J=7.6 Hz, 1H), 6.85 (s,1H), 6.65 (d, J=7.5 Hz, 1H), 3.71 (br s, 2H).

3-(4-methylpiperazin-1-yl)propan-1-amine (1.9 mL, 11 mmoles, 1.1 eq.)was placed in a 3N NaOH aqueous solution (14 mL) and dichloromethane (6mL) was added to the solution. The reaction mixture was cooled down to0° C. with an ice bath and a solution of 3-bromobenzoyl chloride (1.3mL, 10 mmoles, 1 eq.) in dichloromethane (10 mL) was added dropwise. Thereaction mixture was then stirred at room temperature for 10 hours underan inert atmosphere of argon. Upon decantation, the organic phase waswashed with water, dried over MgSO₄, filtered and concentrated underreduced pressure to afford3-bromo-N-(3-(4-methylpiperazin-1-yl)propyl)benzamide (2.8 g, 85%).

¹H NMR (300 MHz, CDCl₃) δ 8.59 (s, 1H), 7.90 (s, 1H), 7.79 (d, J=7.7 Hz,1H), 7.59 (d, J=7.9 Hz, 1H), 7.34-7.24 (m, 1H), 3.60-3.45 (m, 2H),2.60-2.63 (m, 8H), 2.30 (s, 3H), 1.80-1.65 (m, 2H).

According to route (A1), a reaction mixture of3-bromo-N-(3-(4-methylpiperazin-1-yl)propyl)benzamide (169 mg, 0.50mmoles, 1 eq.), (E)-3-(3-(trifluoromethoxy)styryl)aniline (153 mg, 0.55mmoles, 1.1 eq.), Pd₂(dba)₃ (23 mg, 0.025 mmole, 5 mol %), XPhos (24 mg,0.05 mmole, 10 mol %) and K₂CO₃ (276 mg, 2 mmoles, 4 eq.) in t-BuOH (2mL) was heated at 90° C. and stirred for 10 hours under an inertatmosphere of argon. The reaction mixture was then concentrated underreduced pressure and the resulting residue was diluted with ethylacetate. The organic phase was washed with water, dried over MgSO₄,filtered and concentrated under reduced pressure. The resulting residuewas purified by column chromatography on silica gel to give(E)-N-(3-(diethylamino)propyl)-3-((3-(3-(trifluoromethoxy)styryl)phenyl)amino)benzamide (39) (160 mg, 59%).

¹H NMR (300 MHz, CDCl₃) δ 8.19 (s, 1H), 7.57 (s, 1H), 7.42-7.36 (m, 2H),7.33-7.21 (m, 5H), 7.16-7.08 (m, 3H), 7.05-7.01 (m, 3H), 5.89 (s, 1H),3.57 (t, J=5.5 Hz, 2H), 2.66-2.35 (m, 10H), 2.27 (s, 3H), 1.80 (t, J=5.5Hz, 2H).

¹³C NMR (75 MHz, CDCl₃) δ 167.5, 149.9, 143.7, 143.2, 139.6, 138.2,136.5, 130.4, 130.2, 130.0, 129.5, 127.6, 125.1, 120.1, 119.3, 118.9,118.2, 117.1, 116.6, 58.4, 55.1, 53.4, 46.1, 41.0, 24.4.

[M+H]⁺=539

Example 10: Compound (45) in Table I

2-iodobenzoic acid (9.92 g, 40 mmoles, 1 eq.) was placed in toluene (40mL) under an inert atmosphere of argon. Thionyl chloride (6 mL, 80mmoles, 2 eq.) was slowly added. The reaction mixture was heated at 80°C. and stirred for 4 hours. Upon cooling to room temperature, thereaction mixture was concentrated under reduced pressure.N,N-Diethylethylenediamine (3.12 mL, 22 mmoles, 1.1 eq.) was placed in a3N NaOH aqueous solution (15 mL) and dichloromethane (10.7 mL) was addedto the solution. The reaction mixture was cooled down to 0° C. with anice bath and a solution of the 2-iodobenzoyl chloride residue (20mmoles, 4 eq.) in dichloromethane (21.4 mL) was added dropwise. Thereaction mixture was then stirred at room temperature for 18 hours underan inert atmosphere of argon. Upon decantation, the organic phase waswashed with water, dried over MgSO₄, filtered and concentrated underreduced pressure. The resulting residue was purified by columnchromatography on silica gel to afford2-iodo-N-(2-diethylamino-ethyl)benzamide (3.73 g, 51%).

¹H NMR (300 MHz, CDCl₃) δ 7.87 (d, J=7.9 Hz, 1H), 7.42-7.34 (m, 2H),7.13-7.06 (m, 1H), 6.48 (s, 1H), 3.51 (q, J=5.7 Hz, 2H), 2.67 (t, J=5.7Hz, 2H), 2.57 (q, J=7.2 Hz, 4H), 1.02 (t, J=7.2 Hz, 6H).

According to route (A2), a reaction mixture of2-iodo-N-(2-diethylamino-ethyl)benzamide (3.3 g, 9.6 mmoles, 1 eq.),4-(trifluoromethoxy)aniline (1.9 mL, 14.5 mmoles, 1.5 eq.), CuI (183 mg,0.96 mmol, 10 mol %), L-Proline (221 mg, 1.92 mmol, 20 mol %) and K₂CO₃(2.6 g, 19.2 mmoles, 2.0 eq.) in DMSO (10 mL) was heated at 80° C. andstirred for 48 hours under an inert atmosphere of argon. The reactionmixture was then partitioned between ethyl acetate and water. Upondecantation, the aqueous phase was further extracted withdichloromethane. The organic phases were gathered, dried over MgSO₄,filtered and concentrated under reduced pressure. The resulting residuewas purified by column chromatography on silica gel to giveN-(2-diethylamino-ethyl)-2-(4-trifluoromethoxy-phenylamino)-benzamide(45) (1.4 g, 37%).

¹H NMR (300 MHz, CDCl₃) δ 9.55 (s, 1H), 7.48 (d, J=7.7 Hz, 1H),7.30-7.21 (m, 3H), 7.15 (d, J=9.1 Hz, 2H), 7.10 (d, J=9.1 Hz, 2H), 6.78(t, J=7.3 Hz, 1H), 3.44 (t, J=5.9 Hz, 2H), 2.62 (t, J=5.9 Hz, 2H), 2.54(q, J=7.1 Hz, 4H), 1.02 (t, J=7.1 Hz, 6H).

¹³C NMR (75 MHz, CDCl₃) δ 169.5, 145.2, 143.9, 140.8, 132.2, 127.8,122.3, 121.4, 119.1, 118.7, 115.6, 51.3, 46.9, 37.2, 12.1.

[M+H]⁺=396

In a similar manner, compounds (48), (49), (50) and (52) can beprepared.

Example 11: Compound (65) in Table I

4-bromo-3-methylbenzoic acid (9.92 g, 40 mmoles, 1 eq.) was placed intoluene (40 mL) under an inert atmosphere of argon. Thionyl chloride (6mL, 80 mmoles, 2 eq.) was slowly added. The reaction mixture was heatedat 80° C. and stirred for 4 hours. Upon cooling to room temperature, thereaction mixture was concentrated under reduced pressure.3-Methyl-1-butanamine (1.393 mL, 12 mmoles, 2 eq.) was placed in a 3NNaOH aqueous solution (8 mL) and dichloromethane (4 mL) was added to thesolution. The reaction mixture was cooled down to 0° C. with an ice bathand a solution of the 4-bromo-3-methyl benzoyl chloride residue (6mmoles, 1 eq.) in dichloromethane (26 mL) was added dropwise. Thereaction mixture was then stirred at room temperature for 18 hours underan inert atmosphere of argon. Upon decantation, the organic phase waswashed with water, dried over MgSO₄, filtered and concentrated underreduced pressure to afford 4-bromo-3-methyl-N-(3-methylbutyl)benzamide(1.44 g, 84%).

¹H NMR (300 MHz, CDCl₃) δ 7.64 (s, 1H), 7.58 (d, J=8.2 Hz, 1H), 7.40 (d,J=8.2 Hz, 1H), 6.02 (s, 1H), 3.47 (q, J=7.0 Hz, 2H), 2.45 (s, 3H), 1.69(heptuplet, J=6.6 Hz, 1H), 1.51 (q, J=7.0 Hz, 2H), 0.96 (d, J=6.6 Hz,6H).

According to route (A1), a reaction mixture of4-bromo-3-methyl-N-(3-methylbutyl)benzamide (284 mg, 1 mmole, 1 eq.),3-methoxyaniline (185 mg, 1.5 mmole, 1.5 eq.), Pd₂(dba)₃ (46 mg, 0.05mmole, 5 mol %), XPhos (48 mg, 0.10 mmole, 10 mol %) and K₂CO₃ (276 mg,2 mmoles, 2 eq.) in t-BuOH (1 mL) was heated at 100° C. and stirred for22 hours under an inert atmosphere of argon. The reaction mixture wasthen concentrated under reduced pressure and the resulting residue wasdiluted with ethyl acetate. The organic phase was washed with water,dried over MgSO₄, filtered and concentrated under reduced pressure. Theresulting residue was purified by column chromatography on silica gel togive 4-(3-methoxyphenylamino)-3-methyl-N-(3-methylbutyl)benzamide (65)(259 mg, 79%).

¹H NMR (300 MHz, CDCl₃) δ 7.62 (s, 1H), 7.50 (dd, J=1.9, 8.5 Hz, 1H),7.23-7.14 (m, 2H), 6.69-6.59 (m, 2H), 6.54 (dd, J=2.2, 8.1 Hz, 1H), 6.30(d, J=5.4 Hz, 1H), 5.73 (s, 1H), 3.75 (s, 3H), 3.40 (q, J=7.0 Hz, 2H),2.23 (s, 3H), 1.69 (heptuplet, J=6.6 Hz, 1H), 1.50 (q, J=7.0 Hz, 2H),0.93 (d, J=6.6 Hz, 6H). ¹³C NMR (75 MHz, CDCl₃) 167.4, 160.9, 144.7,143.6, 130.4, 130.1, 126.7, 125.8, 125.7, 115.5, 112.2, 107.7, 105.4,55.4, 38.8, 38.5, 26.2, 22.7, 18.0.

[M+H]⁺=327

Example 12: Compound (116) in Table I

N,N-diethyl-N-(2-propynyl)amine (5 g, 45 mmoles, 1.0 eq.) was placed ina dimethylsulfoxide (40.5 mL) and water (4.5 mL) solution, together with1-bromo-3-iodobenzene (12.73 g, 45 mmoles, 1 eq.), NaN₃ (3.51 g, 54mmoles, 1.2 eq.), L-Proline (1.16 g, 9 mmoles, 0.2 eq.), Na₂CO₃ (0.95 g,9 mmoles, 0.2 eq.), sodium ascorbate (3.57 g, 18 mmoles, 0.4 eq.).CuSO₄.5H₂O (2.25 g, 9 mmoles, 0.2 eq.) was added and the reactionmixture was heated at 65° C. and stirred for 16 hours under an inertatmosphere of argon. Upon cooling to room temperature, the reactionmixture was further stirred for 24 hours and then partitioned between aNH₄OH aqueous solution and ethyl acetate. Upon decantation, the aqueousphase was further extracted with ethyl acetate. The organic phases weregathered, washed with a NaCl aqueous solution, dried over MgSO₄,filtered and concentrated under reduced pressure. The resulting residuewas purified by column chromatography on silica gel to afford[1-(3-bromophenyl)-1H-[1,2,3]triazol-4-ylmethyl]diethylamine (9.79 g,70%).

¹H NMR (300 MHz, CDCl₃) δ 7.99-7.90 (m, 1H), 7.71 (d, J=8.1 Hz, 1H),7.56 (d, J=8.1 Hz, 1H), 7.39 (t, J=8.1 Hz, 1H), 3.88 (s, 1H), 2.62 (q,J=7.1 Hz, 1H), 1.13 (t, J=7.1 Hz, 1H).

According to route (E), 4-methoxybenzamide (4.53 g, 30 mmoles, 1 eq.),1-bromo-4-nitrobenzene (6.67 g, 33 mmoles, 1.1 eq.), Pd(OAc)₂ (67.3 mg,0.3 mmoles, 1 mol %), XantPhos (260.4 mg, 0.45 mmoles, 1.5 mol %) andCs₂CO₃ (15.9 g, 45 mmoles, 1.5 eq.) were placed in dioxane (30 mL). Thereaction mixture was heated at 90° C. and stirred for 18 hours under aninert atmosphere of argon. The reaction mixture was then filtered onCelite, washed with CH₂Cl₂ and acetone and the filtrate was concentratedunder reduced pressure. The resulting solid was recrystallized fromethanol and filtered to afford 4-methoxy-N-(4-nitrophenyl)benzamide(4.82 g, 60%).

¹H NMR (300 MHz, CDCl₃) δ 8.27 (d, J=9.3 Hz, 2H), 8.01 (s, 1H), 7.85 (t,J=9.3 Hz, 4H), 7.02 (d, J=9.3 Hz, 2H), 3.90 (s, 3H).

According to route (F), 4-methoxy-N-(4-nitrophenyl)benzamide (4.08 g, 15mmoles, 1 eq.) and 10% Pd/C (750 mg) were placed in EtOH (75 mL). Thereaction mixture was stirred for 6 hours under an atmosphere of H₂. Thereaction mixture was then filtered and the filtrate was concentratedunder reduced pressure to afford N-(4-aminophenyl)-4-methoxybenzamide(2.99 g, 82%).

¹H NMR (300 MHz, CDCl₃) δ 7.87 (d, J=7.9 Hz, 1H), 7.42-7.34 (m, 2H),7.13-7.06 (m, 1H), 6.48 (s, 1H), 3.51 (q, J=5.7 Hz, 2H), 2.67 (t, J=5.7Hz, 2H), 2.57 (q, J=7.2 Hz, 4H), 1.02 (t, J=7.2 Hz, 6H).

According to route (A1), a reaction mixture of[1-(3-bromophenyl)-1H-[1,2,3]triazol-4-ylmethyl]diethylamine (247 mg,0.8 mmole, 1 eq.), N-(4-aminophenyl)-4-methoxybenzamide (213 mg, 0.88mmole, 1.1 eq.), Pd₂(dba)₃ (37 mg, 0.04 mmole, 5 mol %), XPhos (38 mg,0.08 mmole, 10 mol %) and K₂CO₃ (442 mg, 3.2 mmoles, 4 eq.) in t-BuOH (1mL) was heated at 100° C. and stirred for 22 hours under an inertatmosphere of argon. The reaction mixture was then concentrated underreduced pressure and the resulting residue was diluted with ethylacetate. The organic phase was washed with water, dried over MgSO₄,filtered and concentrated under reduced pressure. The resulting residuewas purified by column chromatography on silica gel to giveN-{4-[3-(4-diethylaminomethyl)-[1,2,3]triazol-1-yl)phenylamino]phenyl}-4-methoxybenzamide(116) (196 mg, 52%).

¹H NMR (300 MHz, DMSO-d6) δ 10.03 (s, 1H), 8.60 (s, 1H), 8.45 (s, 1H),7.96 (d, J=8.8 Hz, 2H), 7.70 (d, J=8.8 Hz, 2H), 7.52 (s, 1H), 7.38 (t,J=7.9 Hz, 1H), 7.22 (d, J=7.9 Hz, 1H), 7.16 (d, J=8.8 Hz, 2H), 7.06 (d,J=8.8 Hz, 3H), 3.84 (s, 3H), 3.76 (s, 2H), 2.50-2.41 (m, 4H), 1.03 (t,J=7.1 Hz, 6H). ¹³C NMR (75 MHz, DMSO-d6) δ 164.5, 161.8, 145.8, 137.8,137.7, 133.3, 130.5, 129.5, 127.1, 121.7, 119.1, 114.9, 113.6, 109.8,106.0, 99.5, 55.4, 46.5, 46.1, 11.9.

[M+H]⁺=471

Example 13: Pharmacological Data

13.1. Effects of Compounds on p53 Expression and its TranscriptionalActivity

The compounds of the invention have been tested according to thefollowing pharmacological test and the three compounds have been moreparticularly tested: (40), (1) and (45), all diluted in DMSO. Theireffects were thus compared to cells treated with DMSO. A positivecontrol was used, TG003, a benzothiazole compound known to altersplicing through the inhibition of Clk/Sty family activity (Sigma, ref:T5575).

Two breast cancer cell lines have been treated with 5 μM of drugs for 48hours: wild-type p53 MCF7 cells and mutant p53 (R280K)MDA-MB-231-Luc-D3H2N provided by Caliper Life Science (parental Linesource: American Type Culture collection). The MDA-MB-231-Luc-D3H2Ncells were maintained in two different media during treatment: a DMEMsupplemented with 10% foetal calf serum (FCS) and a FCS-free medium for24 hours followed by 24 hours in a complete medium. The MC7 cells weremaintained in a DMEM/FCS 10% medium.

Cells were harvested after 48 hours of treatment. Total RNAs (RNeasyMini-Kit, Qiagen) and proteins (NuPage LDS 1×, Invitrogen) wereextracted to analyse mRNA expression by nested RT-PCR or RT-qPCR andprotein expression by Western Blot.

1. Protocols:

Analysis of p53 expression was assessed both at mRNA (full-length andtruncated, Δ133p53 isoforms) and protein (full-length) levels by RT-qPCRand Western Blot, respectively.

The p53 isoform cDNA being too long to be specifically quantified byRT-qPCR, a semi-quantitative RT-PCR method was therefore used, requiringhigh quality total RNA 28S/18S ratio (>1.5). After reverse-transcriptionof 500 ng of total RNA using random primers, actin cDNA was amplified byPCR to confirm reverse-transcription efficiency. 0.5 μg of total RNAfrom each tumour sample was reverse-transcribed (AMV RT, 45C, randomprimer) and cDNA quality confirmed by amplification of actin by PCR in30 cycles. p53 isoform cDNA was amplified by 2 nested PCR of 30 cyclesusing primers specific of each isoforms as described in Bourdon et al.(Genes Dev., 2005, 19: 2122). Tumours were considered to express eachp53 isoform after sequencing of the corresponding PCR fragment.

p53 protein expression was determined using a panel of 4 differentantibodies, namely D01, SAPLI, CMI and DO12 as described in Bourdon etal. (Genes Dev., 2005, 19: 2122), to avoid problem of detection due topost-translational modification.

The rabbit anti-p53 antibodies were diluted at 1/1000, and revealed withHorse-Radish Peroxydase (HRP)-conjugated anti-IgG antibodies purchasedfrom GE-Healthcare and diluted at 1/5000 (Ref.: NA9340).

The Western Lightning Chemiluminescence (ECL) reagents were purchasedfrom PerkinElmer (Ref.: NEL103C001EA).

The amount of total proteins in each extract was quantified using BCAkit (promega). 8% SDS-PAGE gels were used. Equal amount (30 μg) ofproteins was loaded on each lane. Proteins were then transferredelectrophoretically on nitrocellulose membrane. Membranes were blockedin TBS/0.1% Tween 20 containing 3% milk for one hour and then incubatedovernight with the primary antibodies diluted in TBS/0.1% Tween 20containing 3% milk. After several washes in TBS/Tween, membranes wereincubated with anti-rabbit Ig antibodies linked to HRP. Membranes weredeveloped with ECL according to the manufacturer's instructions.

Scanned autoradiographs were quantified using AIDA/2D densitometrysoftware.

Analysis of p53 transcriptional activity was assessed through analysisof p53-target gene expression by Western-blot. Four p53-target geneswere studied: Hdm2, Bcl-2, p21 and Bax. Hdm2 is known to bind p53 and tonegatively regulate p53 activity and stability. p21 is known to mediatep53 dependent cycle arrest. Bcl2 is known to have an anti-proliferativeeffect and an apoptotic protective effect, while Bax is known to have apro-apoptic effect. Bax was not detected in both MCF7 andMDA-MB-231-Luc-D3H2N cells, while p21 was not detected inMDA-MB-231-Luc-D3H2N cells maintained in FCS-free medium. TheWestern-blot was performed as above described using the antibodiesdescribed in Bourdon et al., Genes Dev., 2005, 19: 2122.

2. Results

In the wild-type p53 MCF7 cells, the three compounds (40), (1) and (45)did not change total p53 expression (total isoforms, TA) at mRNA levels(FIG. 1A), while compounds (40) and (45) decreased p53 protein levels(full-length) (FIG. 1B, C). This decrease was associated with adecreased expression of Bcl-2 and Hdm2 but not of p21 (FIGS. 1D, E). Thesame pattern of p53-target gene expression was observed with compound(1). Compounds (40) and (1) increased Δ133p53 mRNA level (FIG. 1A).

In mutant p53 MDA-MB-231-Luc-D3H2N cells maintained in normal medium,the three compounds (40), (1) and (45) decreased p53 expression at mRNAlevels (total isoforms, TA) (significant decrease for drug (1), p-value<0.01) (FIG. 2A), while compounds did not change p53 protein levels(full-length) (FIG. 2B, C). Compounds (40) and (1) did not affectp53-target genes expression, while a slight decrease of p53-target geneexpression was observed with compound (45) (FIG. 2D, E).

In the mutant p53 MDA-MB-231-Luc-D3H2N cells maintained in FCS-freemedium, compounds (1) and (45), but not compound (40), decreased p53expression at mRNA levels (total isoforms, TA) (FIG. 3A, blue panel),while compounds did not change p53 protein levels (full-length) (FIG.3B, C). Compound (40) did not affect p53-target genes expression, whilea slight decrease of Hdm2 and of Bcl-2 was observed with compound (1)and (45), respectively (FIG. 3D, E).

3. Conclusion:

Compounds (40) and (45) decreased the expression of both p53 protein(full-length) and its target gene in wild-type p53 MCF7 cells.

Compounds (1) and (45) decreased the p53 mRNA level (total isoforms) inmutant p53 MDA-MB-231-Luc-D3H2N cells maintained in normal medium and inFCS-free medium.

These compounds affect expression of both p53 and its target genes, in ap53 mutation dependent manner.

13.2. Effect of Compounds on p53 Isoforms Expression

1. Protocol:

Expression of p53 isoforms produced by alternative splicing (α, β and γ)was analysed at mRNA levels by RT-qPCR using primers/probe specific ofeach C-terminal variants. The α-forms are the most abundant forms. Inthe low abundant forms, β-forms are more expressed than γ-forms.

A semi-quantitative RT-PCR method requiring high quality total RNA28S/18S ratio (>1.5). After reverse-transcription of 500 ng of total RNAusing random primers, actin cDNA was amplified by PCR to confirmreverse-transcription efficiency. 0.5 μg of total RNA from each tumoursample was reverse-transcribed (AMV RT, 45C, random primer) and cDNAquality confirmed by amplification of actin by PCR in 30 cycles. p53isoform cDNA was amplified by 2 nested PCR of 30 cycles using primersspecific of each isoforms as described by Bourdon et al. (Genes Dev.,2005, 19: 2122). Tumours were considered to express each p53 isoformafter sequencing of the corresponding PCR fragment.

2. Results:

In the wild-type p53 MCF7 cells, the three compounds (40), (1) and (45)did not change α-form expression at mRNA levels (FIG. 4A), while theytend to increase the expression of β- and γ-forms (FIG. 4A).

In the mutant p53 MDA-MB-231-Luc-D3H2N cells maintained in normalmedium, the three compounds (40), (1) and (45) did not change α-formexpression at mRNA levels (FIG. 4B), while they tend to increase theexpression of β- and γ-forms (FIG. 4B). Significant increased of β-forms(by 3-fold) and of γ-forms (by 2-fold) was observed for compound (45).The restoration of γ-forms by compound (1) was confirmed by nestedRT-PCR (FIG. 4B, right panel).

In the mutant p53 MDA-MB-231-Luc-D3H2N cells maintained in FCS-freemedium, the three compounds (40), (1) and (45) did not change α-formexpression at mRNA levels (FIG. 4C), while compound (45) tend toincrease the expression of β- and γ-forms (FIG. 4C). While nosignificant increase of γ-forms was observed by RT-qPCR, restoration ofγ-forms by compound (1) was shown by nested RT-PCR (FIG. 4C, rightpanel).

3. Conclusion:

The tested compounds affect p53 splicing independently of p53 mutationstatus (i.e. increase expression of β- and γ-forms).

13.3. General Conclusion

The compounds of the invention present an effect on p53 isoformsexpression and activity.

As an illustration of said effect, the three here above identifiedtested compounds have different effect on p53 isoforms expression andactivity. They increase the expression of p53β/γ-forms at mRNA levels. Adecrease in p53 protein and its target gene was observed for thecompound (45) (independently of p53 mutation status), for compound (40)(in wild-type p53 cells) and for compound (1) (in mutant p53 cells).

The tested compounds affect p53 family isoforms expression and activity,and consequently the p53 pathway. Surprisingly, the main effect of drugs(40), (1) and (45) is a reduction of p53 expression and its pathway,while, to the knowledge of the inventors, all existing drugs activatep53 pathway. Up to now, to the knowledge of the inventors no drug, whichcan repress p53 expression, has been described.

It follows that the compounds according to the present inventiondemonstrate an effect over the p53 gene isoforms family expression andactivity.

The table below summarizes the effect of the compounds of the inventionon p53 and the therapeutic indications for which they may be proposed.

Therapeutic Effects p53 status (40) (1) (45) indication Decrease ofWild-type X X cancer p53 protein p53 and target Mutant p53 X X Othersthan genes cancer Modification Wild-type X X X cancer of p53 p53isoforms ratio Mutant p53 X X X Others than cancer

13.4 Effect of Drug Compounds on Invasion of MDA-MB231-D3H2LN Cells intoCollagen

Further, some compounds of formula (I) have been tested on invasion inorder to study their activity against cancer as shown below.

Standard operating procedure:

Background:

A key step in the generation of tumor metastasis is the invasion oftumor cells into the extracellular matrix, a major component of which iscollagen. Therefore, the invasion of tumor cells into collagen in vitromay be indicative of the generation of metastasis in vivo. E. g.,MDA-MB231-luc-D3H2LN mouse breast cancer cells display indeed bothhigher invasion into collagen in vitro and a higher metastatic potentialin vivo as compared to MDA-MB231 cells (from which they were derived).Using these MDA-MB231-luc-D3H2LN cells as a model, the aim of theexperiment described here is to identify drug compounds that inhibit theinvasion of tumor cells into collagen in vitro, therefore potentiallyinhibiting also the generation of tumor metastasis in vivo.

Assay Principle:

Step 1: Preparation of cells at the bottom of a collagen gel: Cells aresuspended in a liquid collagen solution (4° C.), distributed intoBSA-coated wells, and then collected at the bottom of the wells bycentrifugation. The collagen is then solidified by incubation at 37° C.The BSA coating improves the adhesion of the collagen gel.

Step 2: Pre-treatment with the compounds to be tested: Concentrated drugsolutions are then added on top of the collagen, and cells arepre-incubated for 24 h with the drugs at low serum conditions (0.025%FBS).

Step 3: Stimulation of invasion: Medium with 5% FBS is then added inorder to stimulate invasion of the cells into the collagen gel.

Step 4: Fixation and staining: Following another 24 h incubation, cellsare fixed and nuclei are stained.

Step 5: Analysis: Finally, plates are analyzed using an automatedmicroscope. Fluorescent beads that have been included into the BSAcoating serve to detect the bottom of the wells. Pictures of the stainednuclei are taken at the same level (0 μm) as well as 25 μm and 50 μmabove.

Note:

In order to detect possible toxic effects, all compounds are tested inparallel in a viability assay. The viability assay is performed inparallel on serum-starved cells (as in the invasion assay) vs. cellsunder normal culture conditions (10% FBS).

Materials:

General equipment: Freezer (−20° C.), refrigerator (4° C.), ice machine,water bath (37° C.), incubator (37° C./5% CO₂), cell culture hood,vortex, vacuum pump, microscope, Malassez cell, Pipet aid, micropipettes(for pipetting 1-1000 μl), multichannel pipettes (for pipetting20-2000), standard cell culture centrifuge, refrigerated centrifuge for96 well plates

General consumables: Sterile 96 well cell culture plates (for theviability assay), sterile tubes (1.5/15/50 ml), sterile pipettes(5/10/25 ml), sterile micropipette tips (for pipetting 1-10000), sterilePasteur pipettes, sterile reagent reservoirs

General products: Sterile PBS, sterile Milli-Q water, DMSO,decomplemented FBS (frozen aliquots), 0.1 N NaOH, 1 M Hepes, MEM withoutserum (not older than 1 month), 2.5×MEM without serum (not older than 1month), MEM with 10% FBS (not older than one month), 0.25% trypsin/1 mMEDTA solution, 37% formaldehyde solution

Specific Equipment:

plate reader: Tecan Infinite F200

automated microscope: Cellomics ArrayScan VTI HCS Reader

Specific Consumables:

sterile black 96 well plates (for the invasion assay): Perkin ElmerViewPlate-96 F TC, ref. 6005225

sterile 96 deep well polypropylene plates (for drug preparation):Starlab, ref. S1896-5110

Specific Products:

rat tail collagen, type 1: BD Biosciences, ref. 354236 (note: each newlot has to be validated)

red fluorescent beads (1 μm diameter): Invitrogen, ref. F13083

Y-27632 (5 mM aqueous solution): Calbiochem, ref. 688001 (in solution)or 688000 (dry powder)

BSA without fatty acids (sterile-filtered 4% aqueous solution): Sigma,ref. A8806 (dry powder)

Hoechst 33342 nuclear stain (10 mg/ml): Invitrogen, ref. H3570

MTS reagent: Promega CellTiter 96® AQueous One Solution Reagent, ref.G3581

drug compounds to be tested: generally 25 or 50 mM in 100% DMSO(aliquots stored at −20° C., then at 4° C. for max. 3 months)

MDA-MB231-luc-D3H2LN cells:

Limits for the cell cultures to be used in the assays:

total passage number: max. 30

last passage: between 2 and 4 days before, between 1:3 and 1:20

cell density: between 50 and 90% (optimally 70%) (between 1 and 2×106cells per 100 mm dish)

Experimental Procedures

General Considerations:

Controls and plate maps:

Invasion assay: Negative control: No drug (just DMSO at equivalentconcentration). Positive control: 10 μM Y-27632. To avoid edge effects,only the 60 central wells B2-G11 are used; lines A and H as well ascolumns 1 and 12 remain free. Each drug is tested at least intriplicate. The positive and negative controls should be tested indouble triplicates at different positions on each plate. Typical platemap (−=negative control, +=positive control, 1-16=16 different drugcompounds):

1 2 3 4 5 6 7 8 9 10 11 12 A B − 1 2 3 4 5 6 7 8 + C − 1 2 3 4 5 6 7 8 +D − 1 2 3 4 5 6 7 8 + E + 9 10 11 12 13 14 15 16 − F + 9 10 11 12 13 1415 16 − G + 9 10 11 12 13 14 15 16 − H

Viability Assays:

No additional controls. The MTS viability assay is based on colorimetricdetection of a product generated by the mitochondrial activity of thecells. Each drug is tested at least in duplicate. To detect potentialdirect interactions with the assay substrate, each drug is also testedin absence of cells (background signals). Typical plate map (controlsand drug compounds as in the invasion assay, lines A-B and E-F: withcells, lines C-D and G-H: without cells; each 1 plate with 10% vs.0.025% FBS):

1 2 3 4 5 6 7 8 9 10 11 12 A − 1 2 3 4 5 6 7 8 + B − 1 2 3 4 5 6 7 8 + C− 1 2 3 4 5 6 7 8 + D − 1 2 3 4 5 6 7 8 + E + 9 10 11 12 13 14 15 16 −F + 9 10 11 12 13 14 15 16 − G + 9 10 11 12 13 14 15 16 − H + 9 10 11 1213 14 15 16 −

The volumes or other quantities indicated in the following are requiredfor testing 16 drug compounds per 96 wells-plate at 5 μM each (+controls) in an invasion assay and each one viability assay onserum-starved cells vs. cells under normal culture conditions accordingto the plate maps above. According to the number of tested compounds,the volumes and other quantities should be adapted for testing more orless compounds or different concentrations.

Day 1: Preparation and Treatment of the Cells (all Steps are PerformedUnder a Cell Culture Hood):

Preparation of 100× concentrated drug solutions in 10% DMSO:

prepare 10% DMSO in sterile PBS: 1.8 ml sterile PBS+0.2 ml DMSO

prepare 100 μl/well 10% DMSO in PBS in 16 wells of a sterile 96 wellpolypropylene plate

add each 1 or 2 μl of the 50 or 25 mM compound stock solutions,respectively

mix by pipetting up and down

Preparation of 4× concentrated drug and control solutions in 0.4% DMSOin MEM+0.1% FBS:

prepare MEM+0.1% FBS: 12 ml MEM without serum+120 FBS (freshly thawedaliquot)

prepare 480 μl/well MEM+0.1% FBS in 20 wells of a sterile 96 deep wellpolypropylene plate

negative controls (no drug): add each 20 μl 10% DMSO in sterile PBS

positive controls (Y-27632): add each 14 μl sterile PBS+20 DMSO+4 μl 5mM Y-27632 (freshly thawed aliquot)

drug compounds: add each 20 μl of the 100× concentrated drug solutionsin 10% DMSO

mix by pipetting up and down

store at RT until use

Coating of the Plates for the Invasion Assay:

mix 9.5 ml MEM without serum+0.5 ml 4% BSA without fatty acids+10vortexed fluorescent beads (i. e. dilute 1:10000), vortex, distribute100 μl/well into the plate for the invasion assay

centrifuge 30′ with 1800×g at 4° C. (e. g. 3000 rpm in a Jouan GR412centrifuge)

remove supernatants by aspiration

Preparation of a 10×106 Cells/Ml Cell Suspension (During theCentrifugation of the Plates):

remove medium, wash cells with ˜10 ml/dish PBS, add 1 ml/dish 0.25%trypsin/1 mM EDTA

incubate 30-60 s at 37° C.

add 5-10 ml/dish pre-warmed MEM+10% FBS

homogenize by pipetting up and down using a 10 ml pipette, pool all

count cells using a Malassez cell

centrifuge 2×106 (or more) cells for 5′ with 150×g at RT (850 rpm in astd. cell culture centrifuge)

remove supernatant, resuspend cell pellet in 0.2 ml (or more,respectively) MEM without serum, yielding 10×106 cells/ml

Preparation of the Invasion Assay (on Ice; Start During theCentrifugation of the Cells):

mix on ice in a pre-chilled tube: example for a 3.4 mg/ml collagen stocksolution; volumes of collagen and water to be adapted according to thestock concentration of each collagen lot:

2.8 ml 2.5×MEM

441 μl water

140 μl 1 M Hepes

49 μl 1 N NaOH

3.5 ml 3.4 mg/ml collagen stock solution (yielding 1.7 mg/ml collagen in7 ml)

homogenize by pipetting gently up and down (keep on ice)

add 700 of the 10×106 cells/ml cell suspension, homogenize by pipettinggently up and down (yields 0.1×106 cells/ml in 1.7 mg/ml collagen in 7ml 1×MEM+20 μM Hepes) (keep on ice)

distribute 100 μl/well (i. e. 10000 cells/well) into the coated wells ofthe plate for the invasion assay (all on ice)

centrifuge 5′ with 200×g at 4° C. (e. g. 1000 rpm in a Jouan GR412centrifuge)

add 200 μl/well PBS to all free wells

incubate 30′ at 37° C./5% CO₂ (solidification of the collagen)

Preparation of the Viability Assay on Serum-Starved Cells:

add 50 μl of the 10×106 cells/ml cell suspension to 5 ml MEM withoutserum (yields 0.1×106 cells/ml)

distribute 100 μl/well of this suspension (i. e. 10000 cells/well) orMEM without serum without cells, respectively, into a standard 96 welltissue culture plate, according to the plate map above

add 200 μl/well PBS to all free wells incubate 30′ at 37° C./5% CO₂

Preparation of the Viability Assay on Cells Under Normal CultureConditions:

add 30 μl of the 10×106 cells/ml cell suspension to 5 ml MEM+10% FBS(yields 0.06×106 cells/ml)

distribute 100 μl/well of this suspension (i. e. 6000 cells/well) orMEM+10% FBS without cells, respectively, into a standard 96 well tissueculture plate, according to the plate map above

add 200 μl/well PBS to all free wells

incubate 30′ at 37° C./5% CO₂

Treatment with the Drugs:

add each 33 μl/well of the 4× concentrated drug solutions in MEM+0.1%FBS to the corresponding wells in all three plates, according to theplate maps above

incubate 24 h at 37° C./5% CO₂

Day 2: Addition of FBS to stimulate the invasion:

Microscopic observation after 24 h of treatment:

examine the cells of the viability assays

Addition of FBS (under a cell culture hood):

prepare MEM+5% FBS: 7.2 ml MEM without serum+0.8 ml FBS (freshly thawedaliquot or rest of the aliquot thawed the day before if kept at 4° C.)

add 33 μl/well to all wells of invasion and viability assays incubate 24h at 37° C./5% CO₂

Day 3: Stop:

Microscopic observation after 48 h of treatment:

examine the cells of the viability assays

Viability assays: MTS assay:

add each 33 μl/well of the MTS reagent, incubate 2.5 h at 37° C./5% CO₂

shake and read absorbance at 490 nm (proportional to the viability)

calculate the background-corrected signals by subtracting the means ofthe background signals in absence of cells from the correspondingsignals in presence of cells

normalize the background-corrected signals with respect to the meansignal of the negative controls (no drug) (viabilities are thusexpressed in “% of control”)

Invasion assays: fixation and staining (formaldehyde must be manipulatedunder a fume cupboard):

freshly prepare 1 μg/ml Hoechst 33342 in 18.5% formaldehyde: 5 ml PBS(not necessarily sterile)+5 ml 37% formaldehyde+1 μl 10 mg/ml Hoechst33342 (note: for one plate, a smaller volume would be sufficient, butthe minimal pipetted volume should not be below 1 μl)

add 50 μl/well to all wells of the invasion assay (yields 4.3%formaldehyde final)

seal with black film (provided with the plates)

incubate at least 7 h at RT

Day 3: 17 (min. 7 h/max. 2 weeks after fixation and staining): Analysisof the invasion assay:

Lecture using the Cellomics ArrayScan VTI HCS Reader:

BioApplication: SpotDetector.V3

Plate type: Perkin Elmer 96 well

Parameters of the Assay Protocol:

objective: 10×(NA 0.45)

apotome: yes (resulting optical slice: 11.7 μM)

fields per well: 8

autofocus in each field

autofocus channel: 1

channel 1 (autofocus on, and photo of the fluorescent beads at thebottom of the wells): filter: XF93-TRITC; exposure time: usually between0.002 and 0.01 s

channel 2 (photo of the stained cells at the same level as thefluorescent beads): filter: XF100-Hoechst; exposure time: usuallybetween 0.02 and 0.1 s; z offset: 0 μM

channel 3 (photo of the stained cells 25 μM above the fluorescentbeads): filter: XF100-Hoechst; exposure time: usually between 0.02 and0.1 s; z offset: −25 μM

channel 4 (photo of the fluorescent cells 50 μM above the fluorescentbeads): filter: XF100-Hoechst; exposure time: usually between 0.02 and0.1 s; z offset: −50 μM

object identification: method: fixed threshold: 100-32767 objectselection parameters: min. max. SpotArea: 20 1000000000000 SpotShapeBFR:0.2 1000 SpotShapeBAR: 0 1000 SpotAvgInten: 200 32767 SpotTotalInten:≤4000 (thus 1000000000000 not limiting) TargetAvgInten: 0 32767TargetTotalInten: 0 1000000000000

Analysis of the Results of the Scan Using vHCS Viewer:

export the results: for each well:

number of valid fields

number of objects in each valid field in each of the channels 2, 3 and 4(“field details”)

mean numbers of objects per valid field for each well, in each of thechannels 2, 3 and 4

exclude wells with less than 6 valid fields per well from furtheranalysis

visually check all photos for any apparent problems, such as badfocusing or obviously inhomogeneous collagen structure (“bubbles”, . . .), . . . ; in case of apparent problems: document, then exclude thecorresponding wells from further analysis

Further Analysis of the Results of the Invasion Assay (Using e. g.Excel):

for each well, calculate the mean invasion distance of the countedcells: (25 μm×number of cells at 25 μm+50 μm×number cells at 50 μm)/sumof cells at 0, 25 and 50 μm

for all four parameters (number of cells at 0 μm, number of cells at 25μm, number of cells at 50 μm, mean invasion distance of the countedcells), calculate means, SD and CV of the replicates (n=6 for thecontrols; n=3 for the samples)

invalidate any replicate with a CV≥50% (compound to be re-tested, orassay to be repeated if CV≥50% for the untreated negative control or thecompound Y-27632-treated positive control). Y27632 is a selectiveinhibitor of the Rho-associated protein kinase p160ROCK of the followingformula

validate the assay only if the mean invasion distance of the cellstreated with 10 μM Y-27632 (positive control) is decreased by ≥40% ascompared to the untreated negative control

plot graphs of all four parameters (number of cells at 0 μm, number ofcells at 25 μm, number of cells at 50 μm, mean invasion distance of thecounted cells)

Results

Anti-invasive effect in MDA-MB231 breast cancer cells: 0.5 fold effectcompared to 10 μM Y-27632 ref. compound.

0.5 fold effect compared Compound to 10 μM Y-27632 (μM) 6 0.7 ± 0.1 91.1 ± 0.4 16 0.75 ± 0.19 17 0.83 ± 0.09 18 0.55 ± 0.05 19 0.25 ± 0.05 210.17 ± 0.04 22  0.3 ± 0.05 24 0.64 ± 0.2  25  0.5 ± 0.21 49 0.13 ± 0.3 50 1   129 0.2

The compounds according to the present invention demonstrate ananti-invasive effect predictive for their activity against cancer.

Therefore, the result of the tests carried out on the compoundsdisclosed in the present invention show that said compounds and theirpharmaceutically acceptable salts, and more particularly compounds (6),(9), (16), (17), (18), (19), (21), (22), (24), (25), (49), (50) and(129) as well as their pharmaceutically acceptable salts, may be used ina method to inhibit, prevent and/or treat cancer.

For this purpose an effective amount of a said compound may beadministered to a patient suffering from cancer.

Others indications than cancer may include inflammation, fibrosis,neurodegenerative diseases, ischemia, atherosclerosis, hepaticdisorders, such as cholestasis, autoimmune diseases, and ethanol-inducedinjuries such as alcoholic liver diseases (ALD) including fatty liver,alcoholic hepatitis and cirrhosis, ribosome biogenesis disorders such asTreacher Collins syndrome (TCS), male infertility, alopecia,neurological defects, endocrinopathy syndrome (ANE syndrome),Shwachman-Diamond syndrome (SDS) and neurofibromatosis type 1 (NF1), andHIV-associated pathologies such as dementia, diabetes and myocardialinfarction.

Therefore, the result of the tests carried out on the compoundsdisclosed in the present invention show that said compounds may beuseful to inhibit, prevent and/or treat cancer and/or atherosclerosisfor patients exhibiting mutated p53, inflammation, fibrosis,neurodegenerative diseases, ischemia, atherosclerosis, pathogenesis of anumber of hepatic disorders, such as cholestasis, autoimmune diseases,and ethanol-induced injuries such as alcoholic liver diseases (ALD)including fatty liver, alcoholic hepatitis and cirrhosis, ribosomebiogenesis disorders such as Treacher Collins syndrome (TCS), maleinfertility, alopecia, neurological defects, endocrinopathy syndrome(ANE syndrome), Shwachman-Diamond syndrome (SDS) and neurofibromatosistype 1 (NF1), and HIV-associated pathologies such as dementia, diabetesand myocardial infarction.

The following type of cancer may more particularly be treated by thecompounds according to the present invention: namely colorectal cancer,pancreatic cancer, lung cancer including non-small cell lung cancer,breast cancer, bladder cancer, gall bladder cancer, thyroid cancer,melanoma, liver cancer, uterine/cervical cancer, oesophageal cancer,kidney cancer, ovarian cancer, prostate cancer, head and neck cancer, orstomach cancer, etc.

For this purpose an effective amount of a said compound may beadministered to a patient suffering from cancer and/or atherosclerosisfor patients exhibiting mutated p53, inflammation, fibrosis,neurodegenerative diseases, ischemia, atherosclerosis, hepaticdisorders, such as cholestasis, autoimmune diseases, ethanol-inducedinjuries such as alcoholic liver diseases (ALD) including fatty liver,alcoholic hepatitis and cirrhosis, ribosome biogenesis disorders such asTreacher Collins syndrome (TCS), male infertility, alopecia,neurological defects, endocrinopathy syndrome (ANE syndrome),Shwachman-Diamond syndrome (SDS) and neurofibromatosis type 1 (NF1), andHIV-associated pathologies such as dementia, diabetes and myocardialinfarction.

The present invention is also related to the use of at least a compoundchosen among a compound of anyone of formula (I), (Iab), (Ia), (Ib),(Ic), (Id), (Ie), (If) as defined above, and compounds (1) to (140) asdefined above, or one of its pharmaceutically acceptable salts accordingto the present invention for the manufacture of a pharmaceuticalcomposition intended for the treatment of cancer and/or atherosclerosisfor patients exhibiting mutated p53, inflammation, fibrosis,neurodegenerative diseases, ischemia, atherosclerosis, hepaticdisorders, such as cholestasis, autoimmune diseases, ethanol-inducedinjuries such as alcoholic liver diseases (ALD) including fatty liver,alcoholic hepatitis and cirrhosis, ribosome biogenesis disorders such asTreacher Collins syndrome (TCS), male infertility, alopecia,neurological defects, endocrinopathy syndrome (ANE syndrome),Shwachman-Diamond syndrome (SDS) and neurofibromatosis type 1 (NF1), andHIV-associated pathologies such as dementia, diabetes and myocardialinfarction.

The present invention also encompasses pharmaceutical compositionscomprising at least a compound chosen among new compounds (9), (16),(17), (18), (19), (20), (21), (22), (23), (24), (25), (37), (38), (39),(48), (49), (50), (52), (83), (84), (85), (86), (87), (88), (89), (90),(91), (92), (123), (124), (125), (126), (127), (128), (129), (130),(131), (132), (133), (134), (135), (136), (137), (138), (139) and (140)as defined above or any pharmaceutically acceptable salt thereof.

Thus, these pharmaceutical compositions contain an effective amount ofsaid compound, and one or more pharmaceutical excipients.

The aforementioned excipients are selected according to the dosage formand the desired mode of administration.

In this context they can be present in any pharmaceutical form which issuitable for enteral or parenteral administration, in association withappropriate excipients, for example in the form of plain or coatedtablets, hard gelatine, soft shell capsules and other capsules,suppositories, or drinkable, such as suspensions, syrups, or injectablesolutions or suspensions, in doses which enable the daily administrationof from 0.1 to 1000 mg of active substance.

The present invention further relates to a method of treatment ofpatients suffering from cancer and/or atherosclerosis said patientsexhibiting mutated p53, inflammation, fibrosis, neurodegenerativediseases, ischemia, atherosclerosis, pathogenesis disorders such ascholestasis, autoimmune diseases and ethanol-induced injuries such asalcoholic liver diseases (ALD) including fatty liver, alcoholichepatitis and cirrhosis, ribosome biogenesis disorders such as TreacherCollins syndrome (TCS), male infertility, alopecia, neurologicaldefects, endocrinopathy syndrome (ANE syndrome), Shwachman-Diamondsyndrome (SDS) and neurofibromatosis type 1 (NF1), and HIV-associatedpathologies such as dementia, diabetes and myocardial infarction forpatients exhibiting a deregulated p53, which comprises at least a stepof administration to a patient suffering thereof of an effective amountof a compound of anyone of formula (I), (Iab), (Ia), (Ib), (Ic), (Id),(Ie) or (If) as defined above and (1) to (140) or one of itspharmaceutically acceptable salts.

1. A compound selected from the group consisting of:

and pharmaceutically acceptable salts thereof.
 2. The compound accordingto claim 1 selected from the group consisting of the compounds (9),(16), (17), (18), (19), (21), (22), (24), (25), (49), (50), (129), andpharmaceutically acceptable salts thereof.
 3. The compound according toclaim 1 selected from the group consisting of the compounds (9), (16),(17), (18), (19), (21), (22), (24), (25), (129), and pharmaceuticallyacceptable salts thereof.
 4. The compound according to claim 1 selectedfrom the group consisting of the compounds (49), (50), andpharmaceutically acceptable salts thereof.
 5. The compound according toclaim 1 selected from the group consisting of the compounds (9), (16),(17), (18), (19), (20), (21), (22), (23), (24), (25), (127), (128),(129), and pharmaceutically acceptable salts thereof.
 6. The compoundaccording to claim 1 selected from the group consisting of the compounds(130), (131), (132), (133), (134), (135), and pharmaceuticallyacceptable salts thereof.
 7. The compound according to claim 1 selectedfrom the group consisting of the compounds (37), (38), (39), (136), andpharmaceutically acceptable salts thereof.
 8. The compound according toclaim 1 selected from the group consisting of the compounds (48), (49),(50), (52), and pharmaceutically acceptable salts thereof.
 9. Thecompound according to claim 1 selected from the group consisting of thecompounds (83), (84), (85), (86), (87), (88), (89), (90), (91), (92),and pharmaceutically acceptable salts thereof.
 10. The compoundaccording to claim 1 selected from the group consisting of the compounds(123), (124), (125) (126), (137), (138), (139), (140), andpharmaceutically acceptable salts thereof.
 11. A pharmaceuticalcomposition comprising at least one compound according to claim 1, orany pharmaceutically acceptable salt thereof.